EP2349479B1 - Elektromagnetische nahinfrarotänderung von stationärem zellmembranpotenzial - Google Patents

Elektromagnetische nahinfrarotänderung von stationärem zellmembranpotenzial Download PDF

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EP2349479B1
EP2349479B1 EP09826549.9A EP09826549A EP2349479B1 EP 2349479 B1 EP2349479 B1 EP 2349479B1 EP 09826549 A EP09826549 A EP 09826549A EP 2349479 B1 EP2349479 B1 EP 2349479B1
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membrane
treatment
nimels
light
energy
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French (fr)
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EP2349479A1 (de
EP2349479A4 (de
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Eric Bornstein
John Wlassich
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Nomir Medical Technologies Inc
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Nomir Medical Technologies Inc
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N5/0613—Apparatus adapted for a specific treatment
    • A61N5/0624—Apparatus adapted for a specific treatment for eliminating microbes, germs, bacteria on or in the body
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N5/0613—Apparatus adapted for a specific treatment
    • A61N5/0616—Skin treatment other than tanning
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B18/00—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body
    • A61B2018/00988—Means for storing information, e.g. calibration constants, or for preventing excessive use, e.g. usage, service life counter
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B18/00—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body
    • A61B18/18—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body by applying electromagnetic radiation, e.g. microwaves
    • A61B18/20—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body by applying electromagnetic radiation, e.g. microwaves using laser
    • A61B2018/2035—Beam shaping or redirecting; Optical components therefor
    • A61B2018/20361—Beam shaping or redirecting; Optical components therefor with redirecting based on sensed condition, e.g. tissue analysis or tissue movement
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B18/00—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body
    • A61B18/18—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body by applying electromagnetic radiation, e.g. microwaves
    • A61B18/20—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body by applying electromagnetic radiation, e.g. microwaves using laser
    • A61B2018/2065—Multiwave; Wavelength mixing, e.g. using four or more wavelengths
    • A61B2018/207—Multiwave; Wavelength mixing, e.g. using four or more wavelengths mixing two wavelengths
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N2005/0626—Monitoring, verifying, controlling systems and methods
    • A61N2005/0627—Dose monitoring systems and methods
    • A61N2005/0628—Dose monitoring systems and methods including a radiation sensor
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N2005/063—Radiation therapy using light comprising light transmitting means, e.g. optical fibres
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N2005/0635—Radiation therapy using light characterised by the body area to be irradiated
    • A61N2005/0643—Applicators, probes irradiating specific body areas in close proximity
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N2005/0635—Radiation therapy using light characterised by the body area to be irradiated
    • A61N2005/0643—Applicators, probes irradiating specific body areas in close proximity
    • A61N2005/0644—Handheld applicators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N2005/0635—Radiation therapy using light characterised by the body area to be irradiated
    • A61N2005/0643—Applicators, probes irradiating specific body areas in close proximity
    • A61N2005/0645—Applicators worn by the patient
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N2005/0658—Radiation therapy using light characterised by the wavelength of light used
    • A61N2005/0659—Radiation therapy using light characterised by the wavelength of light used infrared

Definitions

  • the present invention generally relates to systems for generating infrared optical radiation in selected energies and dosimetries that will modify the bioenergetic steady-state trans-membrane and mitochondrial potentials of irradiated cells through a depolarization effect, and more particularly, relates to systems for membrane depolarization to potentiate antimicrobial and antifungal compounds in target bacterial and/or fungal and/or cancer cells.
  • antibiotics not only prompts generation of resistant bacteria; such as, for example, methicillin-resistant staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE); but also creates favorable conditions for infection with the fungal organisms (mycosis), such as, Candida.
  • MRSA methicillin-resistant staphylococcus aureus
  • VRE vancomycin-resistant enterococci
  • AmB amphotericin B
  • therapies for bacterial and fungal infections include administration of antibacterial and antifungal therapeutics or, in some instances, application of surgical debridement of the infected area. Because antibacterial and antifungal therapies alone are rarely curative, especially in view of newly emergent drug resistant pathogens and the extreme morbidity of highly disfiguring surgical therapies, it has been imperative to develop new strategies to treat or prevent microbial infections.
  • US2004030325 relates to an attachment for a laser emitting device including a body having a first end configured to detachably engage a laser emitting device and a second end, and a cap attached to the second end of the body and having a surface configured to contact the skin of a patient.
  • a control device attached to the body is configured to control the operation of the laser emitting device when the first end of the body is engaged with the laser emitting device.
  • an apparatus for positioning a light delivery head of a therapeutic treatment device in proximity to a body part having a target treatment region.
  • the apparatus includes a positioner including a receptacle defining an at least partially enclosed volume configured to receive at least a portion of the delivery head, the receptacle having a treatment delivery surface including a light transmitting region which is at least partially transparent to therapeutic light from the treatment head, and a light shielding region which is relatively less transparent to the therapeutic light than the light transmitting region; a fixation facility which affixes the receptacle to the body part such that the light transmitting region of the treatment delivery surface is adjacent to the target treatment region; a digital memory; and a communication link configured to selectively couple the memory to the treatment device.
  • the digital memory is readable and writable by the treatment device via the communication link.
  • the digital memory stores information indicative of the identity of the positioner.
  • the information indicative of the identity of the positioner is encrypted.
  • the digital memory stores information indicative of the usage history of the positioner.
  • the information indicative of the usage history of the positioner is encrypted.
  • the apparatus further including at least one sensor, and wherein the communication link configured to selectively couple the at least one sensor to the treatment device.
  • the at least one sensor includes a temperature sensor.
  • the at least one sensor includes at least one selected from the list consisting of: a position sensor, humidity sensor, position sensor, pressure sensor, accelerometer, photodetector, optical power sensor, and optical wavelength sensor.
  • the communication link couples the memory to the treatment device only when the delivery head is received by the receptacle.
  • the communication link includes at least one from the list consisting of: an electrical link, a wired link, a wireless link, a radio link, an optical communication link, and an inductive link.
  • the apparatus further including a connector which connects the delivery head to the receptacle in a desired orientation.
  • the connector is configured to prevent the connection of the delivery head to the receptacle at orientations other than the desired orientation.
  • the apparatus further including at least one indicia for facilitating alignment of the light transmitting region with the target region.
  • the light shielding region is peripheral to the light transmitting region.
  • the fixation facility includes an adhesive material in contact with a portion of the treatment delivery surface.
  • At least a portion of the adhesive material extends beyond the treatment delivery surface.
  • the receptacle includes at least one port providing fluid communication between the at least partially enclosed volume and the exterior of the receptacle.
  • the apparatus further including a microchip which includes the digital memory.
  • the light transmitting region includes a diffusing element which at least partially diffuses therapeutic light from the delivery head to the target region.
  • the light transmitting region is at least partially transparent to light in the near infrared.
  • the apparatus further including the treatment device, and wherein the treatment device includes a controller; the controller is configured to receive information stored in the digital memory via the communication and control the delivery of treatment light based on the information.
  • the information stored in the digital memory includes information indicative of the identity of the positioner or the usage history of the positioner; and the controller is configured to control the delivery of treatment light based on the identity or the usage history of the positioner.
  • the information stored in the digital memory includes information indicative of the usage history of the positioner, and wherein the controller is configured to inhibit delivery of treatment light if any prior use of the positioner is indicated.
  • the information stored in the digital memory includes encrypted information, and the controller is configured to decrypt the encrypted information.
  • the controller is configured to write information to storage in the digital memory via the communication link.
  • the information written to storage in the digital memory includes information indicative of the usage history of the positioner.
  • the treatment device includes: a therapeutic light source configured to generate treatment light; and a delivery assembly which delivers the treatment light to the delivery head; wherein the controller is operatively connected to the light source and controls the light source to provide the treatment light at the target region substantially in a first wavelength range from about 865 nm to about 875 nm or a second radiation range having a wavelength from about 925 nm to about 935 nm, or both wavelength ranges, at a dosimetry including power density of about 0.5 W/cm 2 to about 40 W/cm 2 and an energy density from about 200 J/cm 2 to about 700 J/cm 2 , and a time duration of about 50 to about 720 seconds, and wherein the treatment light is delivered to the target region from the delivery head through the light transmitting region of the treatment delivery surface of the positioner.
  • the controller is operatively connected to the light source and controls the light source to provide the treatment light at the target region substantially in a first wavelength range from 865 nm to 875 nm and a second wavelength range having a wavelength from 925 nm to 935 nm, and at a dosimetry including power density of about 0.5 W/cm 2 to about 5 W/cm 2 and an energy density from about 200 J/cm 2 to about 700 J/cm 2 .
  • the treatment light at the target region has a spot size of at least 1 cm.
  • a NIMELS wavelength includes any wavelength within the ranges of the NIMELS wavelengths described, as well as combinations of such wavelengths.
  • the present invention is directed to systems for reducing the minimum inhibitory concentration (MIC) of antimicrobial molecules (agents) and/or antineoplastic molecules (agents) necessary to attenuate or eliminate microbial and/or neoplastic related pathology, so that the antimicrobial agents that would otherwise be no longer functional at safe human doses will again be useful as adjunctive therapy.
  • MIC minimum inhibitory concentration
  • agents antimicrobial molecules
  • agents antineoplastic molecules
  • neoplastic related pathology so that the antimicrobial agents that would otherwise be no longer functional at safe human doses will again be useful as adjunctive therapy.
  • NIMELS near infrared optical radiation in selected energies and dosimetries
  • NIMELS effect can potentiate existing antimicrobial molecules against microbes infecting and causing harm to human hosts. These effects will render less functional many cellular anabolic reactions (e.g., cell wall formation) and drug-resistance mechanisms (e.g., efflux pumps) that require chemiosmotic electrochemical energy to function.
  • any membrane bound cellular resistance mechanisms or anabolic reaction that makes use of the membrane potential ⁇ , proton motive force ⁇ p, ATP, or the phosphorylation potential ⁇ Gp for their functional energy needs, will be affected by the methods and systems of the present invention.
  • the systems of the present invention utilize optical radiation to potentiate antimicrobial and or antifungal drugs against only targeted undesirable cells (e.g., MRSA or Candida infection in skin) with a selectivity made possible by the fact that mammalian cells are not generally affected by treatments (with molecules or drugs) that are intended to damage the bacterial or fungal cells.
  • targeted undesirable cells e.g., MRSA or Candida infection in skin
  • the applied optical radiation used in accordance with methods and systems of the present invention includes one or more wavelengths ranging from about 800 nm to about 900 nm, at a NIMELS dosimetry, as described herein. In one aspect, wavelengths from about 865 nm to about 885 nm are utilized. In another aspect, such applied radiation has a wavelength from about 905 nm to about 945 nm at a NIMEL dosimetry. In one aspect, such applied optical radiation has a wavelength from about 925 nm to about 935 nm. In a particular aspect, a wavelength of (or narrow wavelength range including) 930 nm can be employed.
  • multiple wavelength ranges include at least two wavelengths in the range of 860 to 940 nm, with particularly preferred embodiments being two wavelengths of about 875 and about 930 nm. It is also preferred that a substantial portion of the power output by the device be in the range of 860 to 940 nm, and more particularly output in the two wavelengths of about 875 and about 930 nm.
  • Microbial pathogens whose bioenergetic systems can be affected by the NIMELS according to the present invention include microorganisms such as, for example, bacteria, fungi, molds, mycoplasms, protozoa, and parasites.
  • the systems of the present invention are used in treating, reducing and/or eliminating the infectious entities known to cause cutaneous or wound infections such as staphyloccocci and enterococci.
  • Staphyloccoccal and enterococcal infections can involve almost any skin surface on the body known to cause skin conditions such as boils, carbuncles, bullous impetigo and scalded skin syndrome.
  • S. aureus is also the cause of staphylococcal food poisoning, enteritis, osteomilitis, toxic shock syndrome, endocarditis, meningitis, pneumonia, cystitis, septicemia and post-operative wound infections.
  • Staphyloccoccal infections can be acquired while a patient is in a hospital or long-term care facility.
  • MRSA methicillin resistant staphylococcus aureus
  • Infections caused by MRSA are frequently resistant to a wide variety of antibiotics (especially ⁇ -lactams) and are associated with significantly higher rates of morbidity and mortality, higher costs, and longer hospital stays than infections caused by non-MRSA microorganisms.
  • Risk factors for MRSA infection in the hospital include colonization of the nares, surgery, prior antibiotic therapy, admission to intensive care, exposure to a MRSA-colonized patient or health care worker, being in the hospital more than 48 hours, and having an indwelling catheter or other medical device that goes through the skin.
  • the systems of the present invention are used in treating, reducing and/or eliminating the infectious entities known as cutaneous Candidiasis.
  • These Candida infections involve the skin, and can occupy almost any skin surface on the body. However, the most often occurrences are in warm, moist, or creased areas (such as armpits and groins). Cutaneous candidiasis is extremely common. Candida is the most common cause of diaper rash, where it takes advantage of the warm moist conditions inside the diaper. The most common fungus to cause these infections is Candida albicans. Candida infection is also very common in individuals with diabetes and in the obese. Candida can also cause infections of the nail, referred to as onychomycosis, infections of the skin surrounding the nail (paronychia) and infections around the comers of the mouth, called angular cheilitis.
  • the term also includes irradiating a cell to increase the sensitivity of the biological contaminant through the lowering of ⁇ with the concomitant generation of ROS of an antimicrobial or antineoplastic agent, wherein the contaminant or tumor is resistant to the agent otherwise. This method can be effected without intolerable risks and/or intolerable side effects on the host subject's tissue other than the biological contaminant.
  • potentiation of an anti-fungal or antibacterial or antineoplastic agent it is meant that the methods and systems of this invention counteract the resistance mechanisms in the fungi, bacteria, or cancer sufficiently for the agent to inhibit the growth and/or proliferation of said fungi, bacteria, or cancer at a lower concentration than in the absence of the present methods and systems.
  • potentiation means that the agent will inhibit the growth and/or proliferation of pathogenic cells thereby treating the disease state at a therapeutically acceptable dosage.
  • microorganism refers to an organism that is microscopic and by definition, too small to be seen by the human eye.
  • microorganisms can be bacteria, fungi, archaea, protists, and the like.
  • the word microbial is defined as pertaining or relating to microorganisms.
  • cell membrane or plasma membrane or mitochondrial membrane
  • lipid bilayer that has a common structure in all living cells. It contains primarily proteins and lipids that are involved in a myriad of important cellular processes.
  • Cell membranes that are the target of the present invention have protein/lipid ratios of >1. Stated another way, none of the target membranes in the contaminent (or moiety, i.e., host tissue) contain greater than 49.99% lipid by dry weight.
  • mitochondria refers to membrane-enclosed organelles, found in most eukaryotic cells (mamallian cells and fungi). Mitochondria are the "cellular power plants,” because they generate most of the eukaryotic cell's supply of ATP, used as a source of chemical energy for the cell.
  • the mitochondria contain inner and outer membranes composed of phospholipid bilayers and proteins. The two membranes, however, have different properties.
  • the outer mitochondrial membrane encloses the entire organelle, has a protein-to-phospholipid ratio similar to the eukaryotic plasma membrane, and the inner mitochondrial membrane forms internal compartments known as cristae and has a protein-to-phospholipid ratio similar to prokaryote plasma membranes. This allows for a larger space for the proteins such as cytochromes to function correctly and efficiently.
  • the electron transport system (“ETS”) is located on the inner mitochondrial membrane. Within the inner mitochondrial membrane are also highly controlled transport proteins that transport metabolites across this membrane.
  • Fluid Mosaic Model refers to a widely held conceptualization of biological membranes as a structurally and functionally asymmetric lipid-bilayer, with a larger variety of embedded proteins that aid in cross-membrane transport.
  • the Fluid Mosaic Model is so named, because the phospholipids shift position in the membrane almost effortlessly (fluid), and because the combination of all the phospholipids, proteins, and glycoproteins present within the membrane give the cell a mosaic image from the outside. This model is based on a careful balance of thermodynamic and functional considerations. Alteration of the membrane thermodynamics affects the function of the membrane.
  • Membrane Dipole Potential ⁇ d (in contrast to the Transmembrane Potential ⁇ ) refers to the potential formed between the highly hydrated lipid heads (hydrophilic) at the membrane surface and the low polar interior of the bilayer (hydrophobic). Lipid bilayers intrinsically possess a substantial Membrane Dipole Potential ⁇ d arising from the structural organization of dipolar groups and molecules, primarily the ester linkages of the phospholipids and water. ⁇ d does not depend upon the ions at the membrane surface and will be used herein to describe five different dipole potentials:
  • Trans-Membrane Potential refers to the electrical potential difference between the aqueous phases separated by a membrane (dimensions mV) and will be given by the symbol ( ⁇ ). ⁇ does depend upon the ions at the membrane surface and will be used herein to describe three different plasma trans-membrane potentials.
  • Mitochondrial Trans-Membrane Potential refers to the electrical potential difference between the compartments separated by the mitochondrial inner membrane (dimensions mV) and will be used herein to describe two different mitochondrial trans-membrane potentials.
  • ⁇ -mito-mam and ⁇ -mito-fungi are important parameters of mitochondrial functionality and give a direct quantitative value to the energy status (redox state) of a cell.
  • mammalian plasma trans-membrane potential refers to the electrical potential difference in the mammalian cell plasma membrane between the aqueous phases.
  • the mammalian plasma membrane potential is different from the bacterial and fungal ⁇ that are primarily generated with H + ions (protons).
  • the major facilitator of the ⁇ is the electrogenic Na + /K + -ATPase pump.
  • ⁇ -plas-mam is generated by the additive qualities of trans-membrane K + diffusion (from the inside to the outside of the cell) and the electrogenic Na + /K + -ATPase pump.
  • Mammalian ATP is generated in the mitochondria via the proton pump.
  • fungal plasma trans-membrane potential ( ⁇ -plas-fungi) refers to the electrical potential difference in the fungal cell plasma membrane.
  • the fungal plasma membrane potential is generated by a membrane-bound H + -ATPase, a high-capacity proton pump that requires ATP to function. This H + -ATPase pump is needed for both fungal growth and stable cell metabolism and maintenance. Fungal ATP is generated in the mitochondria.
  • bacterial plasma trans-membrane potential refers to the electrical potential difference in the bacterial cell plasma membrane.
  • the bacterial plasma membrane potential is generated by the steady-state flow (translocation) of electrons and protons (H + ) across the bacterial plasma membrane that occurs with normal electron transport and oxidative phosphorylation, within the bacterial plasma membrane.
  • a common feature of all electron transport chains is the presence of a proton pump to create a transmembrane proton gradient.
  • bacteria lack mitochondria, aerobic bacteria carry out oxidative phosphorylation (ATP production) by essentially the same process that occurs in eukaryotic mitochondria.
  • P-class ion pump refers to a trans-membrane active transport protein assembly which contains an ATP-binding site (i.e., it needs ATP to function). During the transport process, one of the protein subunits is phosphorylated, and the transported ions are thought to move through the phosphorylated subunit.
  • This class of ion pumps includes the Na + /K + -ATPase pump in the mammalian plasma membrane, which maintains the Na + and K + electrochemical potential ( ⁇ Na + /K + ) and the pH gradients typical of animal cells. Another important member of the P-class ion pumps, transports protons (H + ions) out of and K + ions in to the cell.
  • Na + /K + ATPase refers to a P-class ion pump that is present in the plasma membrane of all animal cells, and couples hydrolysis of one ATP molecule to the export of three Na + ions and the import of two K + ions that maintains the Na + and K + electrochemical potential and the pH gradients typical of animal cells.
  • the inside-negative membrane potential in fungal cells also eukaryotic is generated by transport of H + ions out of the cell by a different ATP powered proton pump.
  • ion exchangers and ion channels refer to transmembrane proteins that are ATP-independent systems, and aid in establishing a plasma membrane potential in mammalian cells.
  • Redox shorthand for reduction/oxidation reaction
  • Redox reactions are chemical reactions in which electrons are transferred from a donor molecule to an acceptor molecule.
  • the term redox comes from the two concepts of reduction and oxidation, and can be explained in the simple terms:
  • redox state describes the redox environment (or level of oxidative stress) of the cells being described.
  • steady-state plasma trans-membrane potential refers to the quantitative Plasma Membrane Potential of a mammalian, fungal or bacterial cell before irradiation in accordance with the methods and systems of the present invention that would continue into the future in the absence of such irradiation.
  • Transient-state plasma membrane potential refers to the Plasma Membrane Potential of a mammalian, fungal or bacterial cell after irradiation in accordance with the methods and systems of the present invention whereby the irradiation has changed the bioenergetics of the plasma membrane.
  • ⁇ -tran will also change the redox state of the cell, as the plasma membrane is where the ETS and cytochromes reside.
  • ⁇ -tran is a state that would not occur without irradiation using methods of the present invention.
  • ⁇ -tran will be used herein to describe three (3) different Transient-state plasma trans-membrane potentials based on species.
  • steady-state mitochondrial membrane potential ( ⁇ -steady-mito) refers to the quantitative Mitochondrial Membrane Potential of mammalian or fungal mitochondria before irradiation in accordance with the methods and systems of the present invention that would continue into the future, in the absence of such irradiation.
  • the steady-state flow of electrons and protons across mitochondrial inner membrane that occurs during normal electron transport and oxidative phosphorylation would be in a steady-state because of a constant flow of conventional redox reactions occurring across the membrane. Any modification of this redox state would cause a transient-state mitochondrial membrane potential.
  • ⁇ -steady-mito will be used herein to describe two (2) different steady-state mitochondrial membrane potentials based on species.
  • transient-state mitochondrial membrane potential ( ⁇ -tran-mito-mam or ⁇ -tran-mito-fungi) refers to the membrane potential of a mammalian or fungal cell after irradiation in accordance with the methods and systems of the present invention whereby the irradiation has changed the bioenergetics of the mitochondrial inner membrane.
  • ⁇ -tran-mito will also change the redox state of the cell, as the inner mitochondrial membrane is where the electron transport system (ETS) and cytochromes reside.
  • ETS electron transport system
  • ⁇ -tran-mito could also drastically affect (the Proton-motive force) ⁇ p-mito-mam and ⁇ p-mito-fungi, as these mitochondrial (H + ) gradients are generated in the mitochondria, to produce adequate ATP for a myriad of cellular functions.
  • ⁇ -tran-mito is a state that would not occur without irradiation in accordance with methods and systems of the present invention.
  • ⁇ -tran-mito will be used herein to describe two (2) different transient-state mitochondrial membrane potentials based on species.
  • cytochrome refers to a membrane-bound hemoprotein that contains heme groups and carries out electron transport.
  • ETS electron transport system
  • cytochromes membrane-associated electron carriers
  • pH Gradient refers to the pH difference between two bulk phases on either side of a membrane.
  • proton electrochemical gradient ( ⁇ H + ) (dimensions kJ mol-1) refers to the electrical and chemical properties across a membrane, particularly proton gradients, and represents a type of cellular potential energy available for work in a cell. This proton electrochemical potential difference between the two sides of a membrane that engage in active transport involving proton pumps, is at times also called a chemiosmotic potential or proton motive force.
  • ⁇ H + is reduced by any means, it is a given that cellular anabolic pathways and resistance mechanisms in the affected cells are inhibited.
  • a pharmacological agent configured and arranged for delivery to the target site (i.e., the co-targeting of an anabolic pathway with ( ⁇ n and Tn) + (pharmacological molecule or molecules)).
  • the term "Ion Electrochemical Gradient ( ⁇ x+)” refers to the electrical and chemical properties across a membrane caused by the concentration gradient of an ion (other than H + ) and represents a type of cellular potential energy available for work in a cell.
  • the Na + ion electrochemical gradient is maintained across the plasma membrane by active transport of Na + out of the cell. This is a different gradient than the proton electrochemical potential, yet is generated from an ATP coupled pump, said ATP produced during oxidative phosphorylation from the mammalian mitochondrial proton-motive force ( ⁇ p-mito-mam).
  • ⁇ X + When ⁇ X + is reduced by any means, it is a given that cellular anabolic pathways and resistance mechanisms in the affected cells are inhibited. This can be accomplished by combining ⁇ n and Tn to irradiate a target site alone, or can be further enhanced with the simultaneous or sequential administration of a pharmacological agent configured and arranged for delivery to the target site (i.e., the co-targeting of an anabolic pathway with ( ⁇ n and Tn) + (pharmacological molecule or molecules)).
  • a pharmacological agent configured and arranged for delivery to the target site (i.e., the co-targeting of an anabolic pathway with ( ⁇ n and Tn) + (pharmacological molecule or molecules)).
  • co-targeting of a bacterial anabolic pathway refers to (the ⁇ n and Tn lowering of ( ⁇ H + ) and/or ( ⁇ x + ) and/or ⁇ cell or mitochondrial membranes) of cells at the target site to affect an anabolic pathway) + (a pharmacological molecule or molecules to affect the same bacterial anabolic pathway) and can refer to any of the following bacterial anabolic pathways that are capable of being inhibited with pharmacological molecules:
  • co-targeting of a fungal anabolic pathway refers to (the ⁇ n and Tn lowering of ( ⁇ H + ) and/or ( ⁇ x + ) of cells at the target site to affect an anabolic pathway) + (a pharmacological agent to affect the same fungal anabolic pathway) and can refer to any of the following fungal anabolic pathways that are capable of being inhibited with pharmacological agents:
  • co-targeting of a cancer anabolic pathway refers to (the ⁇ n and Tn lowering of ( ⁇ H + ) and/or ( ⁇ x+) and/or ⁇ cell or mitochondrial membranes) of cells at the target site to affect an anabolic pathway) + (a pharmacological agent to affect the same cancer anabolic pathway to a greater extent than the non cancerous cells) and can refer to any of the following cancer anabolic pathways that are capable of being inhibited with pharmacological agents:
  • ⁇ p proton-motive force
  • ⁇ the term "proton-motive force" refers to the storing of energy (acting like a kind of battery), as a combination of a proton and voltage gradient across a membrane.
  • the two components of ⁇ p are ⁇ (the transmembrane potential) and ⁇ pH (the chemical gradient of H + ).
  • ⁇ p consists of the H + transmembrane potential ⁇ (negative (acidic) outside) and a transmembrane pH gradient ⁇ pH (alkaline inside).
  • This potential energy stored in the form of an electrochemical gradient is generated by the pumping of hydrogen ions across biological membranes (mitochondrial inner membranes or bacterial and fungal plasma membranes) during chemiosmosis.
  • the ⁇ p can be used for chemical, osmotic, or mechanical work in the cells.
  • the proton gradient is generally used in oxidative phosphorylation to drive ATP synthesis and can be used to drive efflux pumps in bacteria, fungi, or mammalian cells including cancerous cells.
  • ⁇ p-mito-mam refers to the potential energy stored in the form of an (H + ) electrochemical gradient across a mammalian mitochondrial inner membrane. ⁇ p-mito-mam is used in oxidative phosphorylation to drive ATP synthesis in the mammalian mitochondria.
  • ⁇ p-mito-Fungi refers to the potential energy stored in the form of an (H + ) electrochemical gradient across a fungal mitochondrial inner membrane.
  • ⁇ p-mito-Fungi is used in oxidative phosphorylation to drive ATP synthesis in the fungal mitochondria.
  • the term "Fungal Plasma Membrane Proton-motive force ( ⁇ p-plas-Fungi)” refers to the potential energy stored in the form of an (H + ) electrochemical gradient, across a fungal plasma membrane and is generated by the pumping of hydrogen ions across the plasma membrane by a membrane-bound H + -ATPase.
  • This plasma membrane-bound H + -ATPase is a high-capacity proton pump, that requires ATP to function.
  • the ATP for this H + -ATPase is generated from the ⁇ p-mito-Fungi.
  • ⁇ p-plas-Fungi can be used to drive efflux pumps in fungal cells.
  • ⁇ p-plas-Bact Bacterial Plasma Membrane Proton-motive force
  • H + electrochemical gradient
  • ⁇ p-plas-Bact is used in oxidative phosphorylation to drive ATP synthesis in the bacterial plasma membrane and can be used to drive efflux pumps in bacterial cells.
  • anabolic pathway refers to a cellular metabolic pathway that constructs molecules from smaller units. These reactions require energy. Many anabolic pathways and processes are powered by adenosine triphosphate (ATP). These processes can involve the synthesis of simple molecules such as single amino acids and complex molecules such as peptidoglycan, proteins, enzymes, ribosomes, cellular organelles, nucleic acids, DNA, RNA, glucans, chitin, simple fatty acids, complex fatty acids, cholesterols, sterols, and ergosterol.
  • ATP adenosine triphosphate
  • energy transduction refers to proton transfer through the respiratory complexes embedded in a membrane, utilizing electron transfer reactions to pump protons across the membrane and create an electrochemical potential also known as the proton electrochemical gradient.
  • energy transformation in cells refers to chemical bonds being constantly broken and created, to make the exchange and conversion of energy possible. It is generally stated that that transformation of energy from a more to a less concentrated form is the driving force of all biological or chemical processes that are responsible for the respiration of a cells.
  • uncoupler refers to a molecule or device that causes the separation of the energy stored in the proton electrochemical gradient ( ⁇ H + ) of membranes from the synthesis of ATP.
  • uncoupling refers to the use of an uncoupler (a molecule or device) to cause the separation of the energy stored in the proton electrochemical gradient ( ⁇ H + ) of membranes from the synthesis of ATP.
  • ATP adenosine 5'-triphosphate
  • ADP adenosine diphosphate
  • the Gibbs free energy is the energy available ("free") to do work, and the term Gibbs free energy change ( ⁇ G) refers to a change in the free energy available in the membrane to do work
  • This free energy is a function of enthalpy ( ⁇ H), entropy ( ⁇ S), and temperature. (Enthalpy and entropy are discussed below.)
  • phosphorylation potential refers to the ⁇ G for ATP synthesis at any given set of ATP, ADP and Pi concentrations (dimensions: kJ mol -1 ).
  • CCCP refers to carbonyl cyanide m-chlorophenylhydrazone, a highly toxic ionophore and uncoupler of the respiratory chain. CCCP increases the conductance of protons through membranes and acts as a classical uncoupler by uncoupling ATP synthesis from the ⁇ H + and dissipating both the ⁇ and ⁇ pH.
  • depolarization refers to a decrease in the absolute value of a cell's plasma or mitochondrial membrane potential ⁇ . It is a given that depolarization of any bacterial plasma membrane will lead to a loss of ATP and increased free radical formation. It is also a given that mitochondrial depolarization of any eukaryotic cell will lead to a loss of ATP and increased free radical formation.
  • enthalpy change refers to a change in the enthalpy or heat content of a membrane system, and is a quotient or description of the thermodynamic potential of the membrane system.
  • entropy change refers to a change in the entropy of a membrane system to that of a more disordered state at a molecular level.
  • redox stress refers to cellular conditions which differ from the standard reduction/oxidation potential (“redox”) state of the cell. Redox stress includes increased levels of ROS, decreased levels of glutathione and any other circumstances that alter the redox potential of the cell.
  • Reactive Oxygen Species refers to one of the following categories:
  • singlet oxygen refers to (“1O 2 ”) and is formed via an interaction with triplet-excited molecules.
  • Singlet oxygen is a non-radical species with its electrons in antiparallel spins. Because singlet oxygen 1O 2 does not have spin restriction of its electrons, it has a very high oxidizing power and is easily able to attack membranes (e.g., via polyunsaturated fatty acids, or PUFAs) amino acid residues, protein and DNA.
  • membranes e.g., via polyunsaturated fatty acids, or PUFAs
  • energy stress refers to conditions which alter ATP levels in the cell. This could be changes in electron transport and exposure to uncoupling agents or ⁇ altering radiation in mitochondrial and/or plasma membranes.
  • the term "NIMELS effect” refers to the modification of the bioenergetic "state" of irradiated cells at the level of the cell's plasma and mitochondrial membranes from ⁇ -steady to ⁇ -traps with the present invention. Specifically, the NIMELS effect can weaken cellular anabolic pathways or antimicrobial and/or cancer resistance mechanisms that make use of the proton motive force or the chemiosmotic potential for their energy needs.
  • periplasmic space or periplasm refers to the space between the plasma membrane and the outer membrane in gram-negative bacteria and the space between the plasma membrane and the cell wall in gram-positive bacteria and fungi such as the Candida and Trichophyton species.
  • This periplasmic space is involved in various biochemical pathways including nutrient acquisition, synthesis of peptidoglycan, electron transport, and alteration of substances toxic to the cell.
  • periplasmic space is of significant clinical importance as it is where ⁇ -lactamase enzymes inactivate penicillin based antibiotics.
  • the term "efflux pump” refers to an active transport protein assembly which exports molecules from the cytoplasm or periplasm of a cell (such as antibiotics, antifungals, or poisons) for their removal from the cells to the external environment in an energy dependent fashion.
  • the term "effflux pump inhibitor” refers to a compound or electromagnetic radiation delivery system and method which interferes with the ability of an efflux pump to export molecules from a cell.
  • the efflux pump inhibitor of this invention is a form of electromagnetic radiation that will interfere with a pump's ability to excrete therapeutic antibiotics, anti-fungal agents, antineoplastic agents and poisons from cells via a modification of the ⁇ -steady-mam , ⁇ -steady-fungi or, ⁇ -steady-bact.
  • a cell that "utilizes an efflux pump resistance mechanism” it is meant that the bacterial or fungal or cancer cell exports anti-bacterial and/or anti-fungal and/or antineoplastic agents from their cytoplasm or periplasm to the external environment of the cell and thereby reduce the concentration of these agents in the cell to a concentration below what is necessary to inhibit the growth and/or proliferation of the cells.
  • the term “inhibit” means that the rate of growth and/or proliferation of population of cells is decreased, and if possible, stopped.
  • the primary structure refers to the linear arrangement of amino acids; the secondary structure refers to whether the linear amino acid structure forms a helical or ⁇ -pleated sheet structure; tertiary structure of a protein or any other macromolecule is its three-dimensional structure, or stated another way, its spatial organization (including conformation) of the entire single chain molecule; the quaternary structure is the arrangement of multiple tertiary structured protein molecules in a multi-subunit complex.
  • protein stress refers to thermodynamic modification in the tertiary and quaternary structure of proteins, including enzymes and other proteins that participate in membrane transport.
  • the term includes, but is not limited to, denaturation of proteins, misfolding of proteins, cross-linking of proteins, both oxygen-dependent and independent oxidation of inter- and intra- chain bonds, such as disulfide bonds, oxidation of individual amino acids, and the like.
  • pH stress refers to modification of the intracellular pH, i.e., a decrease intracellular pH below about 6.0 or an increase intracellular pH above about 7.5. pH. This may be caused, for example, by exposure of the cell to the invention described herein, and altering cell membrane components or causing changes in the steady-state membrane potential potential ⁇ -steady.
  • anti-fungal molecule refers to a chemical or compound that is fungicidal or fungistatic. Of principle efficacy is the present invention's ability to potentiate anti-fungal molecules by inhibiting anabolic reactions and/or efflux pump activity in resistant fungal strains, or inhibiting other resistance mechanisms that require the proton motive force or chemiosmotic potential for energy.
  • anti-bacterial molecule refers to a chemical or compound that is bacteriacidal or bacteriastatic. Another principal efficacy resides in the present invention's ability to potentiate anti-bacterial molecules by inhibiting efflux pump activity in resistant bacterial strains, or inhibiting anabolic reactions and/or resistance mechanisms that require the proton motive force or chemiosmotic potential for energy.
  • a "sub-inhibitory concentration" of an antibacterial or anti-fungal molecule refers to a concentration that is less than that required to inhibit a majority of the target cells in the population.
  • target cells are those cells that are targeted for treatment including, but not limited to, bacterial, fungi, and cancer cells.
  • a sub-inhibitory concentration refers to a concentration that is less than the Minimum Inhibitory Concentration (MIC), which is defined, unless specifically stated to be otherwise, as the concentration required to produce at least 10% reduction in the growth or proliferation of target cells.
  • MIC Minimum Inhibitory Concentration
  • Minimal Inhibitory Concentration or MIC is defined as the lowest effective or therapeutic concentration that results in inhibition of growth of the microorganism.
  • a therapeutically effective amount of a pharmaceutical agent or molecule refers to a concentration of an agent that, together with NIMELS, will partially or completely relieve one or more of the symptoms caused by the target (pathogenic) cells.
  • a therapeutically effective amount refers to that amount of an agent with NIMELS that: (1) reduces, if not eliminates, the population of target cells in the patient's body, (2) inhibits (i.e., slows, if not stops) proliferation of the target cells in the patients body, (3) inhibits (i.e., slows, if not stops) spread of the infection (4) relieves (if not, eliminates) symptoms associated with the infection.
  • Interaction coefficient is defined as a numerical representation of the magnitude of the bacteriastatic/bacteriacidal and/or fungistatic/fungicidal interaction between the NIMELS laser and/or the antimicrobial molecule, with the target cells.
  • the present invention is directed to perturbing cell membrane biological thermodynamics (bioenergetics) and the consequent diminished capacity of the irradiated cells to adequately undergo normal energy transduction and energy transformation.
  • the methods and systems of the present invention optically alter and modify ⁇ d-plas-mam, ⁇ d-mito-mam, ⁇ d-plas-fungi, ⁇ d-mito-fungi and ⁇ d-plas-bact to set in motion further alterations of ⁇ and ⁇ p in the same membranes. This is caused by the targeted near infrared irradiation of the C-H covalent bonds in the long chain fatty acids of lipid bilayers, causing a variation in the dipole potential ⁇ d.
  • membranes lipid bilayers, see, Figure 1
  • ⁇ d dipole potential arising from the structural association of dipolar groups and molecules, primarily the ester linkages of the phospholipids ( Figure 2 ) and water.
  • dipolar groups are oriented such that the hydrocarbon phase is positive with respect to the outer membrane regions ( Figure 3 ).
  • the degree of the dipole potential is usually large, typically several hundreds of millivolts.
  • the second major potential a separation of charge across the membrane, gives rise to the trans-membrane potential ⁇ .
  • the trans-membrane potential is defined as the electric potential difference between the bulk aqueous phases at the two sides of the membrane and results from the selective transport of charged molecules across the membrane.
  • the potential at the cytoplasm side of cell membranes is negative relative to the extracellular physiological solution ( Figure 4A ).
  • the dipole potential ⁇ d constitutes a large and functionally important part of the electrostatic potential of all plasma and mitochondrial membranes. ⁇ d modifies the electric field inside the membrane, producing a virtual positive charge in the apolar bilayer center. As a result of this "positive charge", lipid membranes exhibit a substantial (e.g., up to six orders of magnitude) difference in the penetration rates between positively and negatively charged hydrophobic ions. ⁇ d also plays an important role in the membrane permeability for lipophilic ions.
  • the energy transduction in biological membranes generally involves three interrelated mechanisms:
  • thermodynamics a state function ( state quantity ), is a property or a system that depends only on the current state of the system. It does not depend on the way in which the system attained its particular state.
  • the present invention facilitates a transition of state in a trans-membrane and/or mitochondrial potential ⁇ , in a temporally dependent manner, to move the bioenergetics of a membrane from a thermodynamic steady-state condition ⁇ -steady to one of energy stress and/or redox stress in a transition state ⁇ -trans.
  • the individual photons of infrared radiation do not contain sufficient energy (e.g., as measured in electron-volts) to induce electronic transitions (in molecules) as is seen with photons of ultraviolet radiation. Because of this, absorption of infrared radiation is limited to compounds with small energy differences in the possible vibrational and rotational states of the molecular bonds.
  • the vibrations or rotations within the lipid bilayer's molecular bonds that absorb the infrared photons must cause a net change in the dipole potential of the membrane. If the frequency (wavelength) of the infrared radiation matches the vibrational frequency of the absorbing molecule (i.e., C-H covalent bonds in long chain fatty acids) then radiation will be absorbed causing a change in ⁇ d. This can happen in ⁇ d-plas-mam, ⁇ d-mito-mam, ⁇ d-plas-fungi, ⁇ d-mito-fungi and ⁇ d-plas-bact. In other words, there can be a direct and targeted change in the enthalpy and entropy ( ⁇ H and ⁇ S) of all cellular lipid bilayers with the methods and systems described herein.
  • ⁇ H and ⁇ S enthalpy and entropy
  • the present invention is based upon a combination of insights that have been introduced above and are derived in part from empirical data, which include the following:
  • the laser system and process of the present invention combine the wavelengths at 5 log less power density than is typically found in a confocal laser microscope such as that used in optical traps ( ⁇ to 500,000 w/cm 2 less power) to advantageously exploit the use of such wavelengths for therapeutic laser systems.
  • cytochrome chains With 870 nm, will additionally alter ⁇ -steady and the redox potential of the membranes that have cytochromes (i.e., bacterial plasma membranes, and fungal and mammalian mitochondria).
  • the NIMELS effect occurs in accordance with methods and systems described herein, importantly, without thermal or ablative mechanical damage to the cell membranes.
  • This combined and targeted low dose approach is a distinct variation and improvement from existing methods that would otherwise cause actual mechanical damage to all membranes within the path of a beam of energy.
  • Entropy in a membrane is a state function whose change in a reaction describes the direction of a reaction due to changes in (energy) heat input or output and the associated molecular rearrangements.
  • the NIMELS effect will modify the entropy "state" of irradiated cells at the level of the lipid bilayer in a temporally dependent manner.
  • This increase in entropy will alter the ⁇ d of all irradiated membranes (mitochondrial and plasma) and hence, thermodynamically alter the "steady-state” flow of electrons and protons across a cell membrane ( Figures 6 and 7 ).
  • This will in turn change the steady-state trans-membrane potential ⁇ -steady to a transient-state membrane potential ( ⁇ -tran). This phenomenon will occur in:
  • Such phenomena can in turn decrease the Gibbs free energy value ⁇ G available for the phosphorylation and synthesis of ATP ( ⁇ Gp).
  • the present invention carries out these phenomena in order to inhibit the necessary energy dependent anabolic reactions, potentiating pharmacological therapies, and/or lowering cellular resistance mechanisms (to antimicrobial, antifungal and antineoplastic molecules) as many of these resistance mechanisms make use of the proton motive force or the chemiosmotic potential for their energy needs, to resist and/or efflux these molecules.
  • the action of chemical uncouplers for oxidative phosphorylation and other bioenergetic work is believed to depend on the energized state of the membrane (plasma or mitochondrial). Further, it is believed that the energized state of the bacterial membrane or eukaryotic mitochondrial inner membrane, is an electrochemical proton gradient ⁇ H + that is established by primary proton translocation events occurring during cellular respiration and electron transport.
  • Agents that directly dissipate (depolarize) the ⁇ H + short-circuits energy coupling, and inhibit bioenergetic work, by inducing a reduction in the membrane potential ⁇ -steady. This will occur while respiration (primary proton translocation) continues apace.
  • the present invention can act as an optical uncoupler by lowering the ⁇ H + and ⁇ p of the following irradiated membranes:
  • Lipid peroxidation is a prevalent cause of biological cell injury and death in both the microbial and mammalian world.
  • strong oxidents cause the breakdown of membrane phospholipids that contain polyunsaturated fatty acids (PUFA's).
  • PUFA's polyunsaturated fatty acids
  • Peroxidation of mitochondrial membranes will have detrimental consequences on the respiratory chains resulting in inadequate production of ATP and collapse of the cellular energy cycle.
  • Peroxidation of the plasma membrane can affect membrane permeability, disfunction of membrane proteins such as porins and efflux pumps, inhibition of signal transduction and improper cellular respiration and ATP formation (i.e., the respiratory chains in prokaryotes are housed in the plasma membranes as prokaryotes do not have mitochondria).
  • a free radical is defined as an atom or molecule that contains an unpaired electron.
  • An example of the damage that a free radical can do in a biological environment is the one-electron (via an existing or generated free radical) removal from bis-allylic C-H bonds of polyunsaturated fatty acids (PUFAs) that will yield a carbon centered free radical.
  • PUFAs polyunsaturated fatty acids
  • a free radical can also add to a nonradical molecule, producing a free radical product. (A* + B ⁇ A-B*) or a nonradical product (A*+B ⁇ A-B)
  • A* + B ⁇ A-B* or a nonradical product (A*+B ⁇ A-B)
  • An example of this would be the hydroxylation of an aromatic compound by *OH.
  • ROS Reactive Oxygen Species
  • Oxygen gas is actually a free radical species. However, because it contains two unpaired electrons in different ⁇ -anti-bonding orbitals that have parallel spin in the ground state, the (spin restriction) rule generally prevents O 2 from receiving a pair of electrons with parallel spins without a catalyst. Consequently O 2 must receive one electron at a time.
  • Superoxide for example, can either act as an oxidizing or a reducing agent.
  • superoxide can reduce cytochrome C. It is generally believed that the reaction rates of superoxide (O 2 - ) with lipids (i.e., membranes) proteins, and DNA are too slow to have biological significance.
  • the protonated form of superoxide hydroperoxyl radical (HOO*) has a lower reduction potential than (O 2 - ), yet is able to remove hydrogen atoms from PUFA's.
  • the pKa value of (HOO * ) is 4.8 and the (acid) microenvironment near biologiocal membranes will favor the formation of hydroperoxyl radicals.
  • the reaction of superoxide (O 2 - ) with any free F e +3 will produce a "perferryl" intermediate which can also react with PUFA's and induce lipid (membrane) peroxidation.
  • Hydrogen peroxide is not a good oxidizing agent (by itself) and cannot remove hydrogen from PUFA's. It can, however, cross biological membranes (rather easily) to exert dangerous and harmful effects in other areas of cells.
  • (H 2 O 2 ) is highly reactive with transition metals inside microcellular environments, (such as Fe +2 and Cu + ) that can then create hydroxyl radicals (*OH) (known as the Fenton Reaction).
  • An hydroxyl radical is one of the most reactive species known in biology.
  • Hydroxyl radicals will react with almost all kinds of biological molecules. It has a very fast reaction rate that is essentially controlled by the hydroxyl radical (*OH) diffusion rate and the presence (or absence) of a molecule to react near the site of (*OH) creation.
  • the standard reduction potential (E0') for hydroxyl radical (*OH) is (+2.31 V) a value that is 7 x greater than (H 2 O 2 ), and is categorized as the most reactive among the biologically relevant free radicals. Hydroxyl radicals will initiate lipid peroxidation in biological membranes, in addition to damaging proteins and DNA.
  • Alkyl peroxyl radicles and alkoxyl radicles are extremely reactive oxygen species and also contribute to the process of propagation of further lipid peroxidation.
  • the altered redox state of irradiated cells and generation of free radicals and ROS because of the ⁇ -steady + ( NIMELS Treatment ) ⁇ ⁇ - trans phenomenon is another object of the present invention. This is an additive effect to further alter cellular bioenergetics and inhibit necessary energy dependent anabolic reactions, potentiate pharmacological therapies, and/or lower cellular resistance mechanisms to antimicrobial, antifungal and antineoplastic molecules.
  • ROS overproduction can damage cellular macromolecules, above all lipids.
  • Lipid oxidation has been shown to modify both the small-scale structural dynamics of biological membranes as well as their more macroscopic lateral organization and altered a packing density dependent reorientation of the component of the dipole moment ⁇ d.
  • Oxidative damage of the acyl chains (in lipids) causes loss of double bonds, chain shortening, and the introduction of hydroperoxy groups. Hence, these changes are believed to affect the structural characteristics and dynamics of lipid bilayers and the dipole potential ⁇ d.
  • Antimicrobial resistance is defined as the ability of a microorganism to survive the effects of an antimicrobial drug or molecule. Antimicrobial resistance can evolve naturally via natural selection, through a random mutation, or through genetic engineering. Also, microbes can transfer resistance genes between one another via mechanisms such as plasmid exchange. If a microorganism carries several resistance genes, it is called multi-resistant or, informally, a "superbug.”
  • Multi-drug resistance in pathogenic bacteria and fungi are a serious problem in the treatment of patients infected with such organisms.
  • it is tremendously expensive and difficult to create or discover new antimicrobial drugs that are safe for human use.
  • there have been resistant mutant organisms that have evolved challenging all known antimicrobial classes and mechanisms. Hence, few antimicrobials have been able to maintain their long-term effectiveness. Most of the mechanisms of antimicrobial drug resistance are known.
  • Staphylococcus aureus ( S. aureus) is one of the major resistant bacterial pathogens currently plaguing civilization. This gram positive bacterium is primarily found on the mucous membranes and skin of close to half of the adult world-wide population. S. aureus is extremely adaptable to pressure from all known classes of antibiotics. S. aureus was the first bacterium in which resistance to penicillin was found in 1947. Since then, almost complete resistance has been found to methicillin and oxacillin. The "superbug" MRSA (methicillin resistant Staphylococcus aureus) was first detected in 1961, and is now ubiquitous in hospitals and communities worldwide. Today, more than half of all S.
  • aureus infections in the United States are resistant to penicillin, methicillin, tetracycline and erythromycin.
  • glycopeptides and oxazolidinones there have been reports of significant resistance (Vancomycin since 1996 and Zyvox since 2003).
  • CA-MRSA community-associated MRSA
  • potentiators and/or inhibitors if not toxic to humans, would be very valuable for the treatment of patients infected with pathogenic and drug-resistant microbes.
  • S. aureus In the United States, as many as 80% of individuals are colonized with S. aureus at some point. Most are colonized only intermittently; 20-30% are persistently colonized. Healthcare workers, persons with diabetes, and patients on dialysis all have higher rates of colonization.
  • the anterior nares are the predominant site of colonization in adults; other potential sites of colonization include the axilla, rectum, and perineum.
  • Daptomycin's mechanism of action involves a calcium-dependent incorporation of the lipopeptide compound into the cytoplasmic membrane of bacteria. On a molecular level, it is calcium binding between two aspartate residues (in the daptomycin molecule) that decreases its net negative charge and permits it to better act with the negatively charged phospholipids that are typically found in the cytoplasmic membrane of gram-positive bacteria. There is generally no interaction with fungi or mammalian cells at therapeutic levels, so it is a very selective molecule.
  • ⁇ p the main component of which is the transmembrane electrical potential gradient ⁇ H +
  • cells cannot make ATP or take up necessary nutrients needed for growth and reproduction.
  • the collapse of ⁇ H + explains the dissimilar (detrimental) effects produced by daptomycin (e.g., inhibition of protein, RNA, DNA, peptidoglycan, lipoteichoic acid, and lipid biosynthesis).
  • daptomycin has a unique mechanism of action, in that it inserts itself into the Gram-positive bacterial plasma membrane, causing a rapid dissolution of membrane potentials ⁇ p. This dissolution of ⁇ p inhibits the macromolecular ATP dependent functions of protein synthesis, DNA replication and peptidoglycan biosynthesis.
  • daptomycin has not been previously described as an efflux pump inhibitor, it does potentiate antibiotics such as gentamycin, where the ⁇ p plays a critical role in drug uptake and antibacterial action.
  • Daptomycin shows synergy with ⁇ -lactam antibiotics that inhibit the ATP dependent function of peptidoglycan biosynthesis.
  • ⁇ -lactams exhibit antibacterial effects outside of the cell membrane, there is no efflux involved with gram-positive resistant species.
  • the phenomenon of daptomycin potentiation with ⁇ -lactams in S. aureus should be a function of lowered macro-molecular synthetic ability in the cytosol to form the peptide precursors of peptidoglycan.
  • This decreased ability for cellular macro-molecular synthesis is most likely could the result of a decreased production of ATP from the dissolution of ⁇ p and perturbed oxidative phosphorylation.
  • This method is suggested, as ATP is consumed in critical reactions constructing elongation of peptide precursors inside the cytosol during the early stages of peptidoglycan synthesis.
  • ⁇ -lactams have no known activity against prokaryotic ribosomes, and studies have shown that daptomycin treatment inhibits the incorporation of the amino acid alanine into a growing peptidoglycan precursor. Similarly, tests on different ⁇ -lactams with daptomycin have revealed considerable synergy in heavily resistant MRSA strains. Also, studies have shown that the minimum inhibitory concentrations of erythromycin, penicillin and tetracycline for wild type S. aureus and daptomycin resistant S. aureus (induced through stepwise selection to increasingly higher levels of daptomycin) are comparable.
  • Fluoroquinolones inhibit the prokaryotic enzymes Topoisomerase II (DNA gyrase) and Topoisomerase IV in Gram-positive and Gram-negative organisms. These interactions prevent the anabolic ATP dependent function of DNA replication and transcription, which are necessary for bacterial survival.
  • fluoroquinolone efflux pump resistance is the result of the energy dependent AcrAB-TolC
  • EmrAB efflux protein complexes is the result of the energy dependent NorA protein complex.
  • Trimethoprim is a dihydrofolate reductase inhibitor that prevents the synthesis of tetrahydrofolic acid, an essential precursor in the de novo synthesis thymidine monophoshpate (dTMP), and hence prevents prokaryotes from synthesizing the nucleotides necessary for DNA replication.
  • Rifampin inhibits bacterial RNA polymerase by binding to the enzymes beta-subunit, and thereby preventing transcription of DNA to RNA. It has been reported that the accumulation of rifampin into S. aureus is unaffected by the metabolic inhibitors CCCP, dinitrophenol (DNP) or reserpine.
  • trimethoprim and rifampin were chosen as "control" antimicrobial molecules in our studies to test against the ⁇ -lactam, macrolide, polyketide and fluoroquinolone antibiotics, for possible potentiation with 870nm/930nm in these studies.
  • Multidrug resistance efflux pumps are now known to be present in gram-positive bacteria, gram-negative bacteria, fungi, and cancer cells. Efflux pumps generally have a poly-specificity of transporters that confers a broad-spectrum of resistance mechanisms. These can strengthen the effects of other mechanisms of antimicrobial resistance such as mutations of the antimicrobial targets or enzymatic modification of the antimicrobial molecules. Active efflux for antimicrobials can be clinically relevant for ⁇ -lactam antimicrobials, marcolides, fluoroquinolones, tetracyclines and other important antibiotics, along with most antifungal compounds including itraconazole and terbinafine.
  • a microbe With efflux pump resistance, a microbe has the capacity to seize an antimicrobial agent or toxic compound and expel it to the exterior (environment) of the cell, thereby reducing the intracellular accumulation of the agent. It is generally considered that the over-expression of one or more of these efflux pumps prevents the intracellular accumulation of the agent to thresholds necessary for its inhibitory activity. Universally in microbes, the efflux of drugs is coupled to the proton motive force that creates electrochemical potentials and/or the energy necessary (ATP) for the needs of these protein pumps. This includes:
  • the approach of the current invention to inhibit efflux pumps is a general modification (optical depolarization) of the membranes ⁇ within the irradiated area, leading to lower electrochemical gradients that will lower the phosphorylation potential ⁇ Gp and energy available for the pumps functional energy needs. It is also the object of the present invention to have the same photobiological mechanism inhibit the many different anabolic and energy driven mechanisms of the target cells, including absorption of nutrients for normal growth.
  • Reserpine inhibits the activity of Bmr and NorA, two gram-positive efflux pumps, by altering the generation of the membrane proton-motive force ⁇ p required for the function of MDR efflux pumps.
  • these molecules are able to inhibit the ABC transporters involved in the extrusion of antibiotics (i.e., tetracycline), the concentrations necessary to block bacterial efflux are neurotoxic in humans.
  • ABC ATP-binding cassette
  • MFS major facilitator superfamily
  • ⁇ -plas-bact During normal cellular metabolism, protons are extruded through the cytoplasmic membrane to form ⁇ -plas-bact. This function also acidifies (lower pH) the narrow region near the bacterial plasma membrane. It has been shown in the gram positive bacterium Bacillus subtilis, that the activities of peptidoglycan autolysins are increased (i.e., no longer inhibited) when the electron transport system was blocked by adding proton conductors. This suggests that ⁇ -plas-bact and ⁇ H + (independent of storing energy for cellular enzymatic functions) potentially has a profound and exploitable influence on cell wall anabolic functions and physiology.
  • ⁇ -plas-bact uncouplers inhibit peptidoglycan formation with the accumulation of the nucleotide precursors involved in peptidoglycan synthesis, and the inhibition of transport of N-acetylglucosamine (GlcNAc), one of the major biopolymers in peptidoglycan.
  • tachyplesin that decreases ⁇ -plas-bact in gram positive and gram negative pathogens.
  • Antimicrobial compositions and pharmaceutical preparations thereof United States Patent 5,610,139 , the entire teaching of which is incorporated herein by reference.
  • This compound was shown at sub-lethal concentrations to have the ability to potentiate the cell wall synthesis inhibitor ⁇ -lactam antibiotic ampicillin in MRSA. It is desirable to couple the multiple influences of an optically lowered ⁇ -plas-bact (i.e., increased cell wall autolysis, inhibited cell wall synthesis, and cell wall antimicrobial potentiation) to any other relevant antimicrobial therapy that targets bacterial cell walls. This is especially relevant in gram positive bacteria such as MRSA that do not have efflux pumps as resistance mechanisms for cell wall inhibitory antimicrobial compounds.
  • ⁇ -mito-fungi is generated in the mitochondria via the electron transport system that then generates ATP via the mitochondrial ATP synthase enzyme system. It is the ATP that then powers the plasma membrane-bound H + -ATPase to generate ⁇ -plas-fungi. It has previously been found that fungal mitochondrial ATP synthase is inhibited by the chemical, polygodial, in a dose-dependent manner ( Lunde and Kubo, Antimicrob Agents Chemother. 2000 July; 44(7): 1943-1953 ).
  • Ergosterol is the structural lipid that is targeted by the majority of relevant commercial antifungal compounds used in medicine today (i.e., azoles, terbinafine and itraconazole).
  • the invention provides a system for modifying the dipole potential ⁇ d of all membranes within the path of a NIMELS beam ( ⁇ d-plas-mam, ⁇ d-mito-mam, ⁇ d-plas-fungi, ⁇ d-mito-fungi, and ⁇ d-plas-bact) to set in motion the cascade of further alterations of ⁇ and ⁇ p in the same membranes.
  • bioenergetic steady-state membrane potentials ⁇ -steady of all irradiated cells are altered to ⁇ -traps values ( ⁇ -trans-mam, ⁇ -trans-fungi, ⁇ -trans-Bact, ⁇ -trans-mito-mam and ⁇ -trans-mito-fungi).
  • such applied optical radiation may have a wavelength from about 850 nm to about 900 nm, at a NIMELS dosimetry, as described herein. In exemplary embodiments, wavelengths from about 865 nm to about 875 nm are utilized. In further embodiments, such applied radiation may have a wavelength from about 905 nm to about 945 nm at a NIMELS dosimetry. In certain embodiments, such applied optical radiation may have a wavelength from about 925 nm to about 935 nm. In representative non-limiting embodiments exemplified hereinafter, the wavelength employed is 930 nm.
  • Bioenergetic steady-state membrane potentials may be modified, in exemplary embodiments, as noted below, and may employ multiple wavelength ranges including ranges bracketing 870 and 930 nm, respectively.
  • NIMELS parameters include the average single or additive output power of the laser diodes and the wavelengths (870 nm and 930 nm) of the diodes. This information, combined with the area of the laser beam or beams (cm 2 ) at the target site, the power output of the laser system and the time of irradiation, provide the set of information which may be used to calculate effective and safe irradiation protocols according to the invention.
  • the near-infrared effect equation takes into account: a) the bacterial response to a photo-damage treatment alone ⁇ growth, neutral or death ⁇ ; b) the bacterial response to an anti-bacterial molecule alone ⁇ growth, neutral or death ⁇ ; and c) the bacterial response to (photo-damage treatment + anti-bacterial molecule ⁇ growth, neutral or death ⁇ ).
  • Np near-infrared potentiation coefficient
  • NIMELS Potentiation Magnitude Scale measures the NIMELS effect number (Ne) between 1 to 10, where the goal is to gain a Ne of ⁇ 4 in reduction of CFU count of a pathogen, at any safe combination of antimicrobial concentration and NIMELS dosimetry.
  • Ne NIMELS effect number
  • CFU count is used here for quantifying pathogenic organism, other means of quantification such as, for example, dye detection methods or polymerase chain reaction (PCR) methods can also be used to obtain values for A, B, and Np parameters.
  • this invention provides methods and systems that will reduced the MIC of antimicrobial molecules necessary to eradicate or at least attenuate microbial pathogens via a depolarization of membranes within the irradiated field which will decrease the membrane potential ⁇ of the irradiated cells.
  • This weakened ⁇ will cause an affiliated weakening of the proton motive force ⁇ p, and the associated bioenergetics of all affected membranes. It is a further object of the present invention that this "NIMELS effect" potentiate existing antimicrobial molecules against microbes infecting and causing harm to human hosts.
  • such applied optical radiation has a wavelength from about 850 nm to about 900 nm, at a NIMELS dosimetry, as described herein. In exemplary embodiments, wavelengths from about 865 nm to about 875 nm are utilized. In further embodiments, such applied radiation has a wavelength from about 905 nm to about 945 nm at a NIMELS dosimetry. In certain embodiments, such applied optical radiation has a wavelength from about 925 nm to about 935 nm. In one aspect, the wavelength employed is 930 nm.
  • irradiation by the wavelength ranges contemplated are performed independently, in sequence, in a blended ratio, or essentially concurrently (all of which can utilize pulsed and/or continuous-wave, CW, operation).
  • NIMELS energy at NIMELS dosimetry to the biological contaminant is applied prior to, subsequent to, or concomitant with the administration of an antimicrobial agent.
  • said NIMELS energy at NIMELS dosimetry can be administered after antimicrobial agent has reached a "peak plasma level" in the infected individual or other mammal. It should be noted that the co-administered antimicrobial agent ought to have antimicrobial activity against any naturally sensitive variants of the resistant target contaminant.
  • the wavelengths irradiated according to the present methods and systems increase the sensitivity of a contaminant to the level of a similar non-resistant contaminant strain at a concentration of the antimicrobial agent of about 0.01 M or less, or about 0.001 M or less, or about 0.0005 M or less.
  • the systems of the invention slow or eliminate the progression of microbial contaminants in a target site, improve at least some symptoms or asymptomatic pathologic conditions associated with the contaminants, and/or increase the sensitivity of the contaminants to an antimicrobial agent.
  • the methods of the invention result in a reduction in the levels of microbial contaminants in a target site and/or potentiate the activity of antimicrobial compounds by increasing the sensitivity of a biological contaminant to an antimicrobial agent to which the biological contaminant has evolved or acquired resistance, without an adverse effect on a biological subject.
  • the reduction in the levels of microbial contaminants can be, for example, at least 10%, 20%, 30%, 50%, 70% or more as compared to pretreatment levels.
  • the sensitivity is potentiated by at least 10%.
  • the invention provides a system to implement the methods according to other aspects of the invention.
  • a system includes a laser oscillator for generating the radiation, a controller for calculating and controlling the dosage of the radiation, and a delivery assembly (system) for transmitting the radiation to the treatment site through an application region.
  • Suitable delivery assemblies/systems include hollow waveguides, fiber optics, and/or free space/beam optical transmission components.
  • Suitable free space/beam optical transmission components include collimating lenses and/or aperture stops.
  • the system utilizes two or more solid state diode lasers to function as a dual wavelength near-infrared optical source.
  • the two or more diode lasers may be located in a single housing with a unified control.
  • the two wavelengths can include emission in two ranges from about 850 nm to about 900 nm and from about 905 nm to about 945 nm.
  • the laser oscillator of the present invention is used to emit a single wavelength (or a peak value, e.g., central wavelength) in one of the ranges disclosed herein. In certain embodiments, such a laser is used to emit radiation substantially within the about 865-875 nm and the about 925-935 nm ranges.
  • Systems according to the present invention can include a suitable optical source for each individual wavelength range desired to be produced.
  • a suitable solid stated laser diode, a variable ultra-short pulse laser oscillator, or an ion-doped (e.g., with a suitable rare earth element) optical fiber or fiber laser is used.
  • a suitable near infrared laser includes titanium-doped sapphire.
  • Other suitable laser sources including those with other types of solid state, liquid, or gas gain (active) media may be used within the scope of the present invention.
  • a therapeutic system includes an optical radiation generation system adapted to generate optical radiation substantially in a first wavelength range from about 850 nm to about 900 nm, a delivery assembly for causing the optical radiation to be transmitted through an application region, and a controller operatively connected to the optical radiation generation device for controlling the dosage of the radiation transmitted through the application region, such that the time integral of the power density and energy density of the transmitted radiation per unit area is below a predetermined threshold. Also within this embodiment, are therapeutic systems especially adapted to generate optical radiation substantially in a first wavelength range from about 865 nm to about 875 nm.
  • a therapeutic system includes an optical radiation generation device that is configured to generate optical radiation substantially in a second wavelength range from about 905 nm to about 945 nm; in certain embodiments the noted first wavelength range is simultaneously or concurrently/sequentially produced by the optical radiation generation device. Also within the scope of this embodiment, are therapeutic systems especially adapted to generate optical radiation substantially in a first wavelength range from about 925 nm to about 935 nm.
  • the therapeutic system can further include a delivery assembly (system) for transmitting the optical radiation in the second wavelength range (and where applicable, the first wavelength range) through an application region, and a controller operatively for controlling the optical radiation generation device to selectively generate radiation substantially in the first wavelength range or substantially in the second wavelength range or any combinations thereof.
  • a delivery assembly for transmitting the optical radiation in the second wavelength range (and where applicable, the first wavelength range) through an application region
  • a controller operatively for controlling the optical radiation generation device to selectively generate radiation substantially in the first wavelength range or substantially in the second wavelength range or any combinations thereof.
  • the delivery assembly comprises one or more optical fibers having an end configured and arranged for insertion in patient tissue at a location within an optical transmission range of the medical device, wherein the radiation is delivered at a NIMELS dosimetry to the tissue surrounding the medical device.
  • the delivery assembly may further comprise a free beam optical system.
  • the controller of the therapeutic system includes a power limiter to control the dosage of the radiation.
  • the controller may further include memory for storing a patient's profile and dosimetry calculator for calculating the dosage needed for a particular target site based on the information input by an operator.
  • the memory may also be used to store information about different types of diseases and the treatment profile, for example, the pattern of the radiation and the dosage of the radiation, associated with a particular application.
  • the optical radiation can be delivered from the therapeutic system to the application site in different patterns.
  • the radiation can be produced and delivered as a continuous wave (CW), or pulsed, or a combination of each. For example, in a single wavelength pattern or in a multi-wavelength (e.g., dual-wavelength) pattern.
  • two wavelengths of radiation can be multiplexed (optically combined) or transmitted simultaneously to the same treatment site.
  • Suitable optical combination techniques can be used, including, but not limited to, the use of polarizing beam splitters (combiners), and/or overlapping of focused outputs from suitable mirrors and/or lenses, or other suitable multiplexing/combining techniques.
  • the radiation can be delivered in an alternating pattern, in which the radiation in two wavelengths are alternatively delivered to the same treatment site.
  • An interval between two or more pulses may be selected as desired according to NIMELS techniques of the present invention.
  • Each treatment may combine any of these modes of transmission.
  • the intensity distributions of the delivered optical radiation can be selected as desired. Exemplary embodiments include top-hat or substantially top-hat (e.g., trapezoidal, etc.) intensity distributions. Other intensity distributions, such as Gaussian may be used.
  • biological contaminant is intended to mean a contaminant that, upon direct or indirect contact with the target site, is capable of undesired and/or deleterious effect(s) on the target site (e.g., an infected tissue or organ of a patient) or upon a mammal in proximity of the target site (e.g., such as, for example, in the case of a cell, tissue, or organ transplanted in a recipient, or in the case of a device used on a patient).
  • Biological contaminants according to the invention are microorganisms such as, for example, bacteria, fungi, molds, mycoplasmas, protozoa, parasites, known to those of skill in the art to generally be found in the target sites.
  • biological contaminants include, but are not limited to, any bacteria, such as, for example, Escherichia, Enterobacter, Bacillus, Campylobacter, Corynebacterium, Klebsiella, Treponema, Vibrio, Streptococcus and Staphylococcus.
  • biological contaminants contemplated include, but are not limited to, any fungus, such as, for example, Trichophyton, Microsporum, Epidermophyton, Candida, Scopulariopsis brevicaulis, Fusarium spp., Aspergillus spp., Alternaria, Acremonium, Scytalidinum dimidiatum, and Scytalidinium hyalinum.
  • Parasites may also be targeted biological contaminants such as Trypanosoma and malarial parasites, including Plasmodium species, as well as molds; mycoplasms and prions.
  • Viruses include, for example, human immuno-deficiency viruses and other retroviruses, herpes viruses, parvoviruses, filoviruses, circoviruses, paramyxoviruses, cytomegaloviruses, hepatitis viruses (including hepatitis B and hepatitis C), pox viruses, toga viruses, Epstein-Barr virus and parvoviruses may also be targeted.
  • the target site to be irradiated need not be already infected with a biological contaminant.
  • the methods of the present invention may be used "prophylactically," prior to infection.
  • Further embodiments include use on medical devices such as catheters, (e.g., IV catheter, central venous line, arterial catheter, peripheral catheter, dialysis catheter, peritoneal dialysis catheter, epidural catheter), artificial joints, stents, external fixator pins, chest tubes, gastronomy feeding tubes, etc.
  • irradiation may be palliative as well as prophylactic.
  • the methods of the invention are used to irradiate a tissue or tissues for a therapeutically effective amount of time for treating or alleviating the symptoms of an infection.
  • the expression "treating or alleviating” means reducing, preventing, and/or reversing the symptoms of the individual treated according to the invention, as compared to the symptoms of an individual receiving no such treatment.
  • the systems according to the invention are used in concomitance with traditional therapeutic approaches available in the art (see, e.g., Goodman and Gilman's, The Pharmacological Basis of Therapeutics, 8th ed, 1990, Pergmon Press, the entire teaching of which is incorporated herein by reference.) to treat an infection by the administration of known antimicrobial agent compositions.
  • antimicrobial composition refers to compounds and combinations thereof that are administered to an animal, including human, and which inhibit the proliferation of a microbial infection (e.g., antibacterial, antifungal, and antiviral).
  • the wide breath of applications contemplated include, for example, a variety of dermatological, podiatric, pediatric, and general medicine to mention but a few.
  • the interaction between a target site being treated and the energy imparted is defined by a number of parameters including: the wavelength(s); the chemical and physical properties of the target site; the power density or irradiance of beam; whether a continuous wave (CW) or pulsed irradiation is being used; the laser beam spot size; the exposure time, energy density, and any change in the physical properties of the target site as a result of laser irradiation with any of these parameters.
  • the physical properties e.g., absorption and scattering coefficients, scattering anisotropy, thermal conductivity, heat capacity, and mechanical strength
  • the target site may also affect the overall effects and outcomes.
  • a biological moiety e.g., a mammalian cell, tissue, or organ
  • NIMELS dosimetry parameters lie between known photochemical and photo-thermal parameters in an area traditionally used for photodynamic therapy in conjunction with exogenous drugs, dyes, and/or chromophores, yet can function in the realm of photodynamic therapy without the need of exogenous drugs, dyes, and/or chromophores.
  • the energy density - also expressible as fluence, or the product (or integral) of particle or radiation flux and time - for medical laser applications in the art typically varies between about 1 J/cm 2 to about 10,000 J/cm 2 (five orders of magnitude), whereas the power density (irradiance) varies from about 1x10 -3 W/cm 2 to over about 10 12 W/cm 2 (15 orders of magnitude).
  • laser exposure duration irradiation time is the primary parameter that determines the nature and safety of laser-tissue interactions.
  • This progression describes a suitable method or basic algorithm that can be used for a NIMELS interaction against a biological contaminant in a tissue.
  • this mathematical relation is a reciprocal correlation to achieve a laser-tissue interaction phenomena.
  • This ratioinale can be used as a basis for dosimetry calculations for the observed antimicrobial phenomenon imparted by NIMELS energies with insertion ofNIMELS experimental data in the energy density and time and power parameters.
  • a practitioner is able to adjust the power density and time to obtain the desired energy density.
  • the threshold energy density needed for a NIMELS interaction with these wavelengths can be maintained independent of the spot-size so long as the desired energies are delivered.
  • the optical energy is delivered through a uniform geometric distribution to the tissues (e.g., a flat-top, or top-hat progression).
  • a suitable NIMELS dosimetry sufficient to generate ROS can be calculated to reach the threshold energy densities required to reduce the level of a biological contaminant and/or to increase the sensitivity of the biological contaminant to an antimicrobial agent that said contaminant is resistant to, but below the level of "denaturization" and "tissue overheating”.
  • NIMELS dosimetries exemplified herein e.g., Onychomycosis
  • NIMELS dosimetries exemplified herein to target microbes in vivo, were from about 200 J/cm 2 to about 700 J/cm 2 for approximately 100 to 700 seconds. These power values do not approach power values associated with photoablative or photothermal (laser/tissue) interactions.
  • the intensity distribution of a collimated laser beam is given by the power density of the beam, and is defined as the ratio of laser output power to the area of the circle in (cm 2 ) and the spatial distribution pattern of the energy.
  • the illumination pattern of a 1.5 cm irradiation spot with an incident Gaussian beam pattern of the area 1.77 cm 2 can produce at least six different power density values within the 1.77 cm 2 irradiation area.
  • These varying power densities increase in intensity (or concentration of power) over the surface area of the spot from 1 (on the outer periphery) to 6 at the center point.
  • a beam pattern is provided which overcomes this inherent error associated with traditional laser beam emissions.
  • Tn is from about 50 to about 300 seconds; in other embodiments, Tn is from about 75 to about 200 seconds; in yet other embodiments, Tn is from about 100 to about 150 seconds. In in vivo embodiments, Tn is from about 100 to about 1200 seconds.
  • NIMEL dosimetry encompasses ranges of power density and/or energy density from a first threshold point at which a subject wavelength according to the invention is capable of optically reducing ⁇ in a target site to a second end-point and/or to increase the sensitivity of the biological contaminant to an antimicrobial agent that said contaminant is resistant to via generation of ROS, immediately before those values at which an intolerable adverse risk or effect is detected (e.g., thermal damage such as poration) on a biological moiety.
  • an intolerable adverse risk or effect e.g., thermal damage such as poration
  • the stopping point contemplated are those at which the adverse effects are considerable and, thus, undesired (e.g., cell death, protein denaturation, DNA damage, morbidity, or mortality).
  • the power density range contemplated herein is from about 0.25 to about 40 W/cm 2 . In other embodiments, the power density range is from about 0.5 W/cm 2 to about 25 W/cm 2 .
  • power density ranges can encompass values from about 0.5 W/cm 2 to about 10 W/cm 2 .
  • Power densities exemplified herein are from about 0.5 W/cm 2 to about 5 W/cm 2 .
  • Power densities in vivo from about 1.5 to about 2.5 W/cm 2 have been shown to be effective for various microbes.
  • Empirical data appears to indicate that higher power density values are generally used when targeting a biological contaminant in an in vitro setting (e.g., plates) rather than in vivo (e.g., toe nail).
  • the energy density range contemplated herein is greater than 50 J/cm 2 but less than about 25,000 J/cm 2 . In other embodiments, the energy density range is from about 750 J/cm 2 to about 7,000 J/cm 2 . In yet other embodiments, the energy density range is from about 1,500 J/cm 2 to about 6,000 J/cm 2 depending on whether the biological contaminant is to be targeted in an in vitro setting (e.g., plates) or in vivo (e.g., toe nail or surrounding a medical device). In certain embodiments (see, in vivo examples below), the energy density is from about 100 J/cm 2 to about 500 J/cm 2 .
  • the energy density is from about 175 J/cm 2 to about 300 J/cm 2 . In yet other embodiments, the energy density is from about 200 J/cm 2 to about 250 J/cm 2 . In some embodiments, the energy density is from about 300 J/cm 2 to about 700 J/cm 2 . In some other embodiments, the energy density is from about 300 J/cm 2 to about 500 J/cm 2 . In yet others, the energy density is from about 300 J/cm 2 to about 450 J/cm 2 .
  • Power densities empirically tested for various in vitro treatment of microbial species were from about 1 W/cm 2 to about 10 W/cm 2 .
  • NIMELS dosimetry values within the power density and energy density ranges contemplated herein for a given circumstance may be empirically done via routine experimentation. Practitioners (e.g., dentists) using near infrared energies in conjunction with periodontal treatment routinely adjust power density and energy density based on the exigencies associated with each given patient (e.g., adjust the parameters as a function of tissue color, tissue architecture, and depth of pathogen invasion).
  • a periodontal infection in a light-colored tissue e.g., a melanine deficient patient
  • a light-colored tissue e.g., a melanine deficient patient
  • the darker tissue will absorb near-infrared energy more efficiently, and hence transform these near-infrared energies to heat in the tissues faster.
  • antibiotic resistant bacteria may be effectively treated according to the methods of the present invention.
  • the methods of this invention may be used to augment traditional approaches, to be used in combination with, in lieu of tradition therapy, or even serially as an effective therapeutic approach. Accordingly, the invention may be combined with antibiotic treatment.
  • antibiotic includes, but is not limited to, ⁇ -lactams, penicillins, and cephalosporins, vancomycins, bacitracins, macrolides (erythromycins), ketolides (telithromycin), lincosamides (clindomycin), chloramphenicols, tetracyclines, aminoglycosides (gentamicins), amphotericns, anilinouracils, cefazolins, clindamycins, mupirocins, sulfonamides and trimethoprim, rifampicins, metronidazoles, quinolones, novobiocins, polymixins, oxazolidinone class (e.g., linezolid), glycylcyclines (e.g., tigecycline), cyclic lipopeptides (e.g., daptomycin), pleuromutilins (e.g.,
  • tetracyclines include, but are not limited to, immunocycline, chlortetracycline, oxytetracycline, demeclocycline, methacycline, doxycycline and minocycline and the like.
  • aminoglycoside antibiotics include, but are not limited to, gentamicin, amikacin and neomycin, and the like.
  • antifungal resistant fungi may be effectively treated according to the methods of the invention.
  • the methods of the present invention may be used to augment traditional approaches, to be used in combination with, in lieu of, traditional therapy, or even serially as an effective therapeutic approach.
  • the invention may be combined with antifungal treatment.
  • antifungal includes, but is not limited to, polyenes, azoles, imidazoles, triazoles, allylamines, echinocandins, cicopirox, flucytosine, griseofulvin, amorolofine, sodarins and combinations thereof (including salts thereof).
  • antineoplastic resistant cancer may be effectively treated according to the methods of the present invention.
  • the methods of the invention may be used to augment traditional approaches, to be used in combination with, in lieu of tradition therapy, or even serially as an effective therapeutic approach. Accordingly, the invention maybe combined with antineoplastic treatment.
  • antiineoplastic includes, but is not limited to, actinomycin, anthracyclines, bleomycin, plicamycin, mitomycin, taxanes, etoposide, teniposide and combinations thereof (including salts thereof).
  • a common tenet in the prior art of trying to find an inhibitor of drug resistance systems in bacteria and fungi, or a potentiator of antimicrobial agents has always been that such agents must be non-toxic to the mammalian tissues that are infected to have any intrinsic value. Furthermore, it has always been a fact that antimicrobials affect bacterial or fungal cellular processes that are not common to the mammalian host, and, hence, are generally safe and therapeutic in nature and design. In the prior art, if antimicrobials, potentiators, and/or resistance reversal entities were to also affect the mammalian cells in the same manner as they damage the pathogens, they could not be used safely as a therapy.
  • cytotoxic wavelengths are employed by the system, which are output at power levels high enough to cause photodamage to microbes without causing substantial photothermal damage to an illuminated target region of a subjects' tissues.
  • the system includes an optical radiation generation device, which is configured and arranged to generate near infrared optical radiation (i) substantially in: a first wavelength range from about 865 nm to less than 880 nm, and preferably 875 nm and/or a second wavelength range from about 925 nm to about 935 nm, or in both wavelength ranges.
  • the dosimetry includes a power density of about 0.5 W/cm2 to about 5 W/cm2 and an energy density from about 200 J/cm2 to about 700 J/cm2 at the illuminated target region.
  • the time duration is about 50 to about 720 seconds.
  • the dosimetry is sufficient to produce photodamage in the biological contaminant without causing substantial photothermal or photomechanical damage to biological tissue of the subject at the illuminated target region.
  • the system includes a delivery assembly for causing the optical radiation to be transmitted to illuminate the target region of the subject, wherein substantially all of the near infrared optical radiation transmitted from the optical radiation generation device to the target region by the delivery assembly is in the first wavelength range or the second wavelength range, or both, and wherein the near infrared optical radiation preferably has a spot size at the target region of at least 1.0 cm.
  • the system further includes a controller operatively connected to the optical radiation generation device for controlling dosage of the near infrared optical radiation transmitted to the target region of the subject at the dosimetry sufficient to produce photodamage in the biological contaminant without causing substantial photothermal or photomechanical damage to biological tissue of the subject at the illuminated target region.
  • the controller is operatively connected to the optical radiation generation device for controlling dosage of the radiation transmitted to the target region at the dosimetry sufficient to produce photodamage in the biological contaminant without causing substantial photothermal or photomechanical damage to biological tissue of the subject at the illuminated target region.
  • the delivery assembly is configured and arranged to simultaneously deliver the optical radiation to a plurality of target regions, wherein at each target region the optical radiation has a dosimetry sufficient to produce photodamage in the biological contaminant without intolerable adverse effects on biological tissue at the target region.
  • the experimental data supports a universal alteration of ⁇ and ⁇ p among all cell types, and hence leads to the notion that not only the electromechanical, but also the electro-dynamical aspects of all cell membranes, have no differing properties that can adequately be separated. This indicates that all cells in the path of the beam are affected with depolarization, not only the pathogenic cells (non-desired cells).
  • Table 2 MIC values for Susceptible, Intermediate and Resistant S . aureus Minimum Inhibitory Concentration (MIC) Interpretive Standards ( ⁇ g/ml) for Staphylococcus sp. Antimicrobial Agent Susceptible Intermediate Resistant Penicillin ⁇ 0.12 - ⁇ 0.25 Methicillin ⁇ 8 - ⁇ 16 Aminoglycosides Gentamicin ⁇ 4 8 ⁇ 16 Tanamycin ⁇ 16 32 ⁇ 64 Macrolides Erythromycin ⁇ 0.5 1-4 ⁇ 8 Tetracycline Tetracycline ⁇ 4 8 ⁇ 16 Fluoroquinolone Ciprofloxacin ⁇ 1 2 >4 Folate Pathway Inhibitors Trimethoprim ⁇ 8 - ⁇ 16 Ansamycins Rifampin ⁇ 1 2 ⁇ 4
  • C. albicans ATCC 14053 liquid cultures were grown in YM medium (21g/L, Difco) medium at 37°C.
  • a standardized suspension was aliquoted into selected wells in a 24-well tissue culture plate. Following laser treatments, 100 ⁇ L was removed from each well and serially diluted to 1:1000 resulting in a final dilution of 1:5x10 6 of initial culture. An aliquot of each final dilution were spread onto separate plates. The plates were then incubated at 37°C for approximately 16-20 hours. Manual colony counts were performed and recorded. Table 4.
  • C. albicans ATCC 14053 liquid cultures were grown in YM medium (21g/L, Difco) medium at 37°C.
  • a standardized suspension was aliquoted into selected wells in a 24 -well tissue culture plate. Following laser treatments each lased and control sample were treated as per directions of individual assay.
  • the following parameters illustrate the general bacterial methods according to the invention as applied to HEK293 cells for the in vitro experiments.
  • HEK293 cells were seeded into appropriate wells of a 24-well plate at a density of 1 x 10 5 cells/ml (0.7ml total volume) in Freestyle medium (Invitrogen). Cells were incubated in a humidified incubator at 37 °C in 8% CO 2 for approximately 48 hours prior to the experiment. Cells were approximately 90% confluent at the time of the experiment equating to roughly 3 x 10 5 total cells. Immediately prior to treatment, cells were washed in pre-warmed phosphate buffer saline (PBS) and overlaid with 2 ml of PBS during treatment.
  • PBS pre-warmed phosphate buffer saline
  • fluorescent dyes that can be taken up by intact cells and accumulate within the intact cells within 15 to 30 minutes without appreciable staining of other protoplasmic constituents. These dye indicators of membrane potential have been available for many years and have been employed to study cell physiology. The fluorescence intensity of these dyes can be easily monitored, as their spectral fluorescent properties are responsive to changes in the value of the trans-membrane potentials ⁇ -steady.
  • These dyes generally operate by a potential-dependent partitioning between the extracellular medium and either the membrane or the cytoplasm of membranes. This occurs by redistribution of the dye via interaction of the voltage potential with an ionic charge on the dye. This fluorescence can be eliminated in about 5 minutes by the protonophore carbonyl cyanide m -chlorophenylhydrazone (CCCP), indicating that maintenance of dye concentration is dependent on the inside-negative transmembrane potential maintained by functional ETS and ⁇ p.
  • CCCP protonophore carbonyl cyanide m -chlorophenylhydrazone
  • the data indicates that the fluorescence of cells is dissipated (less than control of unirradiated or "unlased” cells) by pre-treatment (of the cells) with the NIMELS laser system, indicating that the NIMEL laser interacted with respiratory processes and oxidative phosphorylation of the cells via the plasma membranes.
  • ⁇ 1 - ⁇ 2 0 Will uphold that the addition sub-lethal NIMEL irradiation on the cell culture has no effect on ⁇ -steady.
  • BacLightTM Bacterial Membrane Potential Kit (B34950, Invitrogen U.S.).
  • the Bac LightTM Bacterial Membrane Potential Kit provides of carbocyanine dye DiOC2(3) (3,3'-diethyloxacarbocyanine iodide, Component A) and CCCP (carbonyl cyanide 3-chlorophenylhydrazone, Component B), both in DMSO, and a 1 x PBS solution (Component C).
  • DiOC2(3) exhibits green fluorescence in all bacterial cells, but the fluorescence shifts toward red emission as the dye molecules self associate at the higher cytosolic concentrations caused by larger membrane potentials.
  • Proton ionophores such as CCCP destroy membrane potential by eliminating the proton gradient, hence causing higher green fluorescence.
  • Green fluorescence emission was calculated using population mean fluorescence intensities for control and lased samples at sub-lethal dosimetry: Table 6.
  • MRSA Dosimetry Progression First lasing procedure Both 870 and 930 Second lasing procedure 930 alone Parameters Output Power (W) Beam Spot (cm) Area of Spot(cm2) Time (sec) 870 at 4.25 W and 930 at 4.25 W for 16 min followed by 8.5 1.5 1.77 960 930 at 8.5W for 7 min 8.5 1.5 1.77 420
  • Second lasing procedure 930 alone Parameters Output Power (W) Beam Spot (cm) Area of Spot (cm2) Time (sec) Laser #1 Test (H-1) 870 at 4 W and 930 at 4 W for 18 min followed by 8.0 1.5 1.77 1080 Test (H-1) 930 at 8W for 8 min 8.0 1.5 1.77 480 Laser #2 Test (H-2) 870 at 4.25 Wand 930 at 4.25 W for 18 min 8.5 1.5 1.77 1080 followed by Test (H-2) 930 at 8.5 W for 8 min 8.5 1.5 1.77 480 Laser #3 Test (H-3) 870 at 4 W and 930 at 4 W for 20 min followed by 8.0 1.5 1.77 1200 Test (H-3) 930 at 8 W for 10 min 8.0 1.5 1.77 600
  • Red/green ratios were calculated using population mean fluorescence intensities for control and lased samples at sub-lethal dosimetry:
  • the data shows that the fluorescence of mitochondria is dissipated (less than control unlased cells) by pre-treatment (of the cells) with the NIMELS laser system, the results indicate that the NIMELS laser interacted with respiratory processes and oxidative phosphorylation of the cells in mitochondria of fungal and mammalian cells.
  • ⁇ 1 - ⁇ 2 0 Will uphold that the addition sub-lethal NIMEL irradiation on the cell culture mitochondria has no effect on ⁇ -steady-mito.
  • the loss of mitochondrial membrane potential ( ⁇ ) is a hallmark for apoptosis.
  • the APO LOGIX JC-1 Assay Kit measures the mitochondrial membrane potential in cells.
  • JC-1 (5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide) exists as a monomer in the cytosol (green) and also accumulates as aggregates in the mitochondria which stain red. Whereas, in apoptotic and necrotic cells, JC-1 exists in monomeric form and stains the cytosol green. Table 8.
  • the (APO LOGIX JC-1) kit measures membrane potential by conversion of green fluorescence to red fluorescence.
  • Figure 10A the appearance of red color has been measured and plotted, which should only occur in cells with intact membranes, and the ratio of green to red is shown in Figure 10B for both control and lased samples.
  • Figure 10B the ratio of green to red is shown in Figure 10B for both control and lased samples.
  • the red fluorescence is reduced in the lased sample while the ratio of green to red increases, indicating depolarization.
  • the data shows that the fluorescence of mitochondria is dissipated (less than control unlased cells) by pre-treatment (of the cells) with the NIMELS laser system, the results indicate that the NIMELS laser interacted with respiratory processes and oxidative phosphorylation of the cells in mitochondria of mammalian cells.
  • ⁇ 1 - ⁇ 2 0 Will uphold that the addition sub-lethal NIMEL irradiation on the mammalian cell culture mitochondria has no effect on ⁇ -steady-mito-mam.
  • Mitochondrial Membrane Potential Detection Kit (APO LOGIX JC-1) (Cell Technology Inc., 950 Rengstorff Ave, Suite D; Mountain View CA 94043).
  • the loss of mitochondrial membrane potential ( ⁇ ) is a hallmark for apoptosis.
  • the APO LOGIX JC-1 Assay Kit measures the mitochondrial membrane potential in cells.
  • JC-1 (5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenz-imidazolylcarbocyanine iodide) exists as a monomer in the cytosol (green) and also accumulates as aggregates in the mitochondria which stain red.
  • JC-1 exists in monomeric form and stains the cytosol green.
  • Table 9 Mamallian Cell Dosimetries First lasing procedure : Both 870 and 930 Second lasing procedure 930 alone Parameters Output Power (W) Beam Spot (cm) Area of Spot(cm2) Time (sec) Test (H-2) 870 at 4.25 W and 930 at 4.25 W for 18 min followed by 8.5 1.5 1.77 1080 Test (H-2) 930 at 8.5 W for 10 mm 8.5 1.5 1.77 600
  • the (APO LOGIX JC-1) kit measures membrane potential by conversion of green fluorescence to red fluorescence.
  • Figure 11A the appearance of red color has been measured and plotted, which should only occur in cells with intact membranes, and the ratio of green to red is shown in Figure 11B for both control and lased samples.
  • ROS reactive oxygen species
  • Glutathione is the most abundant thiol (SH) compound in animal tissues, plant tissues, bacteria and yeast. GSH plays many different roles such as protection against reactive oxygen species and maintenance of protein SH groups. During these reactions, GSH is converted into glutathione disulfide (GSSG: oxidized form of GSH). Since GSSG is enzymatically reduced by glutathione reductase, GSH is the dominant form in organisms.
  • DTNB (5,5'-Dithiobis(2-nitrobenzoic acid)), known as Ellman's Reagent, was developed for the detection of thiol compounds.
  • ROS glutathione antioxidant system
  • Erythromycin has been used for people with allergies to penicillins. Erythromycin's mechanism of action is to prevent growth and replication of bacteria by obstructing bacterial protein synthesis. This is accomplished because erythromycin binds to the 23S rRNA molecule in the 50S of the bacterial ribosome, thereby blocking the exit of the growing peptide chain thus inhibiting the translocation of peptides. Erythromycin resistance (as with other marcolides) is rampant, wide spread, and is accomplished via two significant resistance systems:
  • Trimethoprim is an antibiotic that has historically been used in the treatment of urinary tract infections. It is a member of the class of antimicrobials known as dihydrofolate reductase inhibitors. Trimethoprim's mechanism of action is to interfere with the system of bacterial dihydrofolate reductase (DHFR), because it is an analog of dihydrofolic acid. This causes competitive inhibition of DHFR due to a 1000 fold higher affinity for the enzyme than the natural substrate.
  • DHFR bacterial dihydrofolate reductase
  • trimethoprim inhibits synthesis of the molecule tetrahydrofolic acid.
  • Tetrahydrofolic acid is an essential precursor in the de novo synthesis of the DNA nucleotide thymidylate.
  • Bacteria are incapable of taking up folic acid from the environment (i.e., the infection host) and are thus dependent on their own de novo synthesis of tetrahydrofolic acid. Inhibition of the enzyme ultimately prevents DNA replication.
  • Trimethoprim resistance generally results from the overproduction of the normal chromosomal DHFR, or drug resistant DHFR enzymes. Reports of trimethoprim resistance S . aureus have indicated that the resistance is chromosomally of the mediated type or is encoded on large plasmids. Some strains have been reported to exhibit both chromosomal and plasmid-mediated trimethoprim resistance. In the gram positive pathogen S. aureus, resistance to trimethoprim is due to genetic mutation, and there have been no reports that trimethoprim is actively effluxed out of cells.
  • a major route of drug resistance in bacteria and fungi is the active export (efflux) of antibiotics out of the cells such that a therapeutic concentration in not obtained in the cytoplasm of the cell.
  • Active efflux of antibiotics is mediated by a series of transmembrane proteins in the cytoplasmic membrane of gram positive bacteria and the outer membranes of gram negative bacteria.
  • antibiotic resistance that is mediated via efflux pumps, is most relevant in gram positive bacteria for marcolides, tetracyclines and fluoroquinolones.
  • ⁇ -lactam efflux mediated resistance is also of high clinical relevance.
  • ⁇ 1 - ⁇ 3 0 Will uphold that the addition of erythromycin produces no deleterious effect after sub-lethal NIMEL irradiation, on normal growth of MRSA colonies.
  • EXPERIMENTAL CONTROL (no laser) trimeth erythro trimeth erythro AGAR 2 ug/ml 4 ug/ml AGAR 2 ug/ml 4 ug/ml B-4 1 84 110 39 B-4 1 180 213 196 B-4 2 88 125 35 B-4 2 230 198 168 B-4 3 120 138 39 B-4 3 241 240 175 B-4 4 114 115 28 B-4 4 220 220 177 B-4 5 117 100 27 B-4 5 smeared 145 195
  • Tetracycline is considered a bacteriostatic antibiotic, meaning that it hampers the growth of bacteria by inhibiting protein synthesis. Tetracycline accomplishes this by inhibiting action of the bacterial 30S ribosome through the binding of the enzyme aminoacyl-tRNA. Tetracycline resistance is often due to the acquisition of new genes, which code for energy-dependent efflux of tetracyclines, or for a protein that protects bacterial ribosomes from the action of tetracyclines.
  • Rifampin is a bacterial RNA polymerase inhibitor, and functions by directly blocking the elongation of RNA.
  • Rifampicin is typically used to treat mycobacterial infections, but also plays a role in the treatment of methicillin-resistant Staphylococcus aureus (MRSA) in combination with fusidic acid, a bacteriostatic protein synthesis inhibitor. There are no reports of rifampin resistance via efflux pumps in MRSA.
  • AGAR 90ug/ml 4ug/ml AGAR 90 ug/ml 4ug/ml E1-1 307 210 42 E1-1 270 183 240 E1-2 300 200 56 E1-2 210 210 256 E1-3 300 280 46 E1-3 224 166 268 E1-4 310 378 48 E1-4 semared 228 310 E1-5 250 280 42 E1-5 215 188 255 E2-1 246 272 18 E2-1 240 274 280 E2-2 254 320 28 E2-2 310 210 283 E2-3 174 330 27 E2-3 190 180 263 E2-4 170 semared 16 E2-4 257 240 260 E2-5 240 284 18 E2-5 275 310 E3-1 310 270 72 E3-1 280 288 368 E3-2 280 225 67 E3-2 320 280 380 E3-3 260 284 45 E3-3 310 210 375 E3-4 210 200 47 E3-4 320 290 390 E3-5 220
  • Methicillin is a ⁇ -lactam that was previously used to treat infections caused by gram-positive bacteria, particularly ⁇ -lactamase-producing organisms such as S. aureus that would otherwise be resistant to most penicillins, but is no longer clinically used.
  • MRSA methicillin-resistant S. aureus
  • methicillin acts by inhibiting the synthesis of peptidoglycan (bacterial cell walls).
  • ⁇ -plas-bact uncouplers inhibit peptidoglycan formation with the accumulation of the nucleotide precursors involved in peptidoglycan synthesis, and the inhibition of transport of N-acetylglucosamine (GlcNAc), one of the major biopolymers in peptidoglycan.
  • Bacitracin will potentiate the multiple influences of an optically lowered ⁇ -plas-bact on a growing cell wall (i.e., increased cell wall autolysis, inhibited cell wall synthesis). This is especially relevant in gram positive bacteria such as MRSA, that do not have efflux pumps as resistance mechanisms for cell wall inhibitory antimicrobial compounds.
  • the NIMELS laser and its concomitant optical ⁇ -plas-bact lowering phenomenon is synergistic with cell wall inhibitory antimicrobial in MRSA. Without wishing to be bound by theory, this must function via the inhibition of anabolic (periplasmic) ATP coupled functions, as MRSA does not have efflux pumps for methicillin.
  • Bacitracin is a mixture of cyclic polypeptides produced by Bacillus subtilis. As a toxic and difficult-to-use antibiotic, bacitracin cannot generally be used orally, but used topically.
  • Bacitracin interferes with the dephosphorylation of the C 55 -isoprenyl pyrophosphate, a molecule which carries the building blocks of the peptidoglycan bacterial cell wall outside of the inner membrane in gram negative organisms and the plasma membrane in gram positive organism.
  • ⁇ -plas-bact uncouplers inhibit peptidoglycan formation with the accumulation of the nucleotide precursors involved in peptidoglycan synthesis, and the inhibition of transport of N-acetylglucosamine (GlcNAc), one of the major biopolymers in peptidoglycan.
  • Bacitracin potentiates the multiple influences of an optically lowered ⁇ -plas-bact on a growing cell wall (i.e., increased cell wall autolysis, inhibited cell wall synthesis). This is especially relevant in gram positive bacteria such as MRSA, that do not have efflux pumps as resistance mechanisms for cell wall inhibitory antimicrobial compounds.
  • Ergosterol is the structural lipid that is targeted by the majority of relevant commercial antifungal compounds used in medicine today (i.e., azoles, terbinafine and itraconazole) including lamisil and sporanox (and generic counterparts thereof).
  • Lamisil (like other allylamines) inhibits ergosterol synthesis by inhibiting squalene expoxidase, an enzyme that is part of the fungal cell wall synthesis pathway.
  • the NIMELS laser at sub-lethal dosimetry on C . albicans potentiates lamisil and sporanox due to of an optically lowered ⁇ -plas-fungi and/or ⁇ -mito-fungi by depolarizing the membranes and depleting cellular ATP in the fungus.
  • the data indicates that the addition of the antifungal lamisil and/or sporanox after sub-lethal irradiation, results in the reduction in growth of these C . albicans colonies, as follows: ⁇ 1 - ⁇ 2 0 Will uphold that the addition of Sporanox produces no deleterious effect after sub-lethal NIMEL irradiation, on normal growth of C . albicans colonies. ⁇ 1 - ⁇ 2 > 0 Will uphold that the addition of Sporanox produces a deleterious effect after sub-lethal NIMEL irradiation, on normal growth of C . albicans colonies.
  • microorganisms exemplified include E. coli K-12, multi-drug resistant E. coli, Staphylococcus aureus, methicillin-resistant S . aureus, Candida albicans, and Trichophyton rubrum.
  • NIMELS parameters include the average single or additive output power of the laser diodes, and the wavelengths (870 nm and 930 nm) of the diodes. This information, combined with the area of the laser beam or beams (cm 2 ) at the target site, provide the initial set of information which may be used to calculate effective and safe irradiation protocols according to the invention.
  • the power density of a given laser measures the potential effect of NIMELS at the target site. Power density is a function of any given laser output power and beam area, and may be calculated with the following equations:
  • Total energy distribution may be measured as energy density (Joules/cm 2 ). As discussed infra, for a given wavelength of light, energy density is the most important factor in determining the tissue reaction.
  • Treatment Time seconds Energy Density Joules / cm 2 Output power Density W / cm 2
  • Treatment Time seconds Energy Joules Laser Output Power Watts
  • the therapeutic system may also include a computer database storing all researched treatment possibilities and dosimetries.
  • the computer (a dosimetry and parameter calculator) in the controller is preprogrammed with algorithms based on the above-described formulas, so that any operator can easily retrieve the data and parameters on the screen, and input additional necessary data (such as: spot size, total energy desired, time and pulse width of each wavelength, tissue being irradiated, bacteria being irradiated) along with any other necessary information, so that any and all algorithms and calculations necessary for favorable treatment outcomes can be generated by the dosimetry and parameter calculator and hence run the laser.
  • the bacterial kill rate (as measured by counting Colony Forming Units or CFU on post-treatment culture plates) ranged from 93.7% (multi-drug resistant E. coli ) to 100% (all other bacteria and fungi).
  • E. coli K12 liquid cultures were grown in Luria Bertani (LB) medium (25 g/L). Plates contained 35 mL of LB plate medium (25 g/L LB, 15 g/L bacteriological agar). Culture dilutions were performed using PBS. All protocols and manipulations were performed using sterile techniques.
  • LB Luria Bertani
  • Liquid cultures of E. coli K12 were set up as described previously. An aliquot of 100 ⁇ L was removed from the subculture and serially diluted to 1:1200 in PBS. This dilution was allowed to incubate at room temperature approximately 2 hours or until no further increase in O.D. 600 was observed in order to ensure that the cells in the PBS suspension would reach a static state (growth) with no significant doubling and a relatively consistent number of cells could be aliquoted further for testing.
  • a standardized suspension was aliquoted into selected wells in a 24-well tissue culture plate. Following laser treatments, 100/ ⁇ L was removed from each well and serially diluted to 1:1000 resulting in a final dilution of 1:5x105 of initial culture. 3x100 ⁇ L of each final dilution were spread onto separate plates. The plates were then incubated at 37°C for approximately 16-20 hours. Manual colony counts were performed and recorded. A digital photograph of each plate was also taken.
  • T. rubrum ATCC 52022 liquid cultures were grown in peptone-dextrose (PD) medium at 37 °C.
  • PD peptone-dextrose
  • a standardized suspension was aliquoted into selected wells in a 24 -well tissue culture plate. Following laser treatments, aliquots were removed from each well and spread onto separate plates. The plates were then incubated at 37 °C for approximately 91 hours. Manual colony counts were performed and recorded after 66 hours and 91 hours of incubation. While control wells all grew the organism, 100% of laser-treated wells as described herein had no growth. A digital photograph of each plate was also taken.
  • This synergistic ability is significant to human tissue safety, as the 930 nm optical energy, heats up a system at a greater rate than the 870 nm optical energy, and it is beneficial to a mammalian system to produce the least amount of heat possible during treatment.
  • This synergistic effect is significant to human tissue safety, as the 930 nm optical energy, heats up a system at a greater rate than the 870 nm optical energy, and it is beneficial to a mammalian system to produce the least amount of heat possible during treatment.
  • This simultaneous synergistic ability is significant to human tissue safety, as the 930 nm optical energy, heats up a system at a greater rate than the 870 nm optical energy, and it is beneficial to a mammalian system to produce the least amount of heat possible during treatment.
  • Experimental in vitro data also demonstrates that when applied at safe thermal dosimetries, there is less additive effect with the 830 nm wavelength, and the NIMELS 930 nm wavelength when they are used simultaneously, In fact, Experimental in vitro data demonstrates that 17% less total energy, 17% less energy density, and 17% less power density is required to achieve 100 % E. coli antibacterial efficacy when 870 nm is combined simultaneously with 930 nm vs. the commercially available 830 nm. This, again, substantially reduces heat and harm to an in vivo system being treated with the NIMELS wavelengths. Table 26. E.
  • NIMELS wavelengths 870 nm and 930 nm
  • FIG. 17 shows: (a) the synergistic effects ofNIMELS with methicillin, penicillin and erythromycin in growth inhibition of MRSA colonies; data show that penicillin and methicillin is being potentiated by sub-lethal NIMELS dosimetry by inhibiting the Bacterial Plasma Membrane Proton-motive force ( ⁇ p-plas-Bact ) thereby inhibiting peptidoglycan synthesis anabolic processes that are co-targeted with the drug; and (b) that erythromycin is potentiated to a greater extent, because the Nimek effect is inhibiting the Bacterial Plasma Membrane Proton-motive force ( ⁇ p-plas-Bact ) that supplies the energy for protein synthesis anabolic processes and erythromycin resistance efflux pumps.
  • the treated and a control untreated suspension were diluted and plated in triplicate on trypic soy agar with or without 30 ⁇ g/ml methicillin. After 24hrs of growth at 37 °C colonies were counted.
  • CFU colony forming units
  • Conditions D1 through H1 showed a similar reduction in CFU on the methicillin plates in treated and untreated samples.
  • Conditions A1, B1 and C1 showed 30%, 33%, or 67% fewer CFU in the laser treated samples compared to the untreated controls, respectively.
  • the treated and a control untreated suspension were diluted and plated in triplicate on trypic soy agar with or without 30 ⁇ g/ml methicillin. After 24hrs of growth at 37 °C colonies were counted.
  • MRSA Dosimetry Progression 11-09-06 Experiment #3 MRSA Dosimetry Progression 11-09-06 First lasing procedure : Both 870 and 930 Second lasing procedure 930 alone Parameters Output Power (W) Beam Spot (cm) Area of Spot (cm2) Time (sec) Total Energy Joules Energy Density (J/cm 2 ) Power Density (W/cm 2 ) Temp Initial C Temp Final C Test (1) 870 at 5.5W and 930 at 5.5W for 12 min followed by 11.0 1.5 1.77 720 7920 4482 6.22 22.0 48.1 Test (1) 930 at 8W for 6 min 8.0 1.5 1.77 360 2880 1630 4.53 48.1 47.7 Test (2) 870 at 5.5W and 930 at 5.5W for 12 min followed by 11.0 1.5 1.77 720 7920 4482 6.22 22.9 48.8 Test (2) 930 at 8W for 6 min 8.0 1.5 1.
  • the treated and a control untreated suspension were diluted and plated in pentuplicate on trypic soy agar with or without 30 ⁇ g/ml methicillin. After 24hrs of growth at 37 °C colonies were counted.
  • the treated and a control untreated suspension were diluted and plated in pentuplicate on trypic soy agar with or without 30 ⁇ g/ml methicillin (Groups A4 and B4), 0.5 ⁇ g/ml penicillin G (Groups C4 and D4) or 4 ⁇ g/ml erythromycin (Groups E4 and F4).
  • the inventor performed a dosimetry titration on himself to ascertain the safe, maximum level of energy and time of exposure that could be delivered to human dermal tissue without burning or otherwise damaging the irradiated tissues.
  • Time/Temperature assessments were charted to ensure the thermal safety of these laser energies on human dermal tissues (data not shown).
  • he exposed his great toe to both 870 nm and 930 nm for up to 233 seconds, while measuring toenail surface temperature with a laser infrared thermometer. He found that using the above dosimetries, at a surface temperature of 37.5°C, with 870 nm and 930 nm together with a combined Power Density of 1.70 W/cm 2 , pain resulted and the laser was turned off.
  • the dosimetry that was used for the treatment of the first subject was the same as that used during the inventor's self-exposure (shown above).
  • the temperature parameters on the surface of the nail also were equivalent to the temperatures found by the inventor on self-exposure.
  • the treated toes showed significantly reduced Tinea pedis and scaling surrounding the nail beds, which indicated a decontamination of the nail plate that was acting as a reservoir for the fungus.
  • the control nails were scraped with a cross-cut tissue bur, and the shavings were saved to be plated on mycological media.
  • the treated nails were scraped and plated in the exact same manner.
  • Treated plates did not show any growth whereas untreated control culture plates showed significant growth.
  • the first subject was followed for 120 days, and received four treatments under the same protocol.
  • Figure 18 shows a comparison of the pretreatment (A), 60 days post-treatment (B), 80 days post-treatment (C), and 120 days post-treatment (D) toenails.
  • healthy and non-infected nail plate was covering 50% of the nail area and growing from healthy cuticle after 120 days.
  • Ne near-infrared effect
  • Example XXI Laser Treatment for Microbial Reduction and Elimination of Nasal Colonization of MRSA
  • the Nomir Near Infrared Microbial Elimination Laser System (NOVEONTM Model 1120 dual-wavelength diode laser was employed for this study.
  • the laser operates in continuous wave format at two wavelengths, 870 nm (+/- 5 nm) and 930 nm (+/- 5 nm).
  • This device is a class II non-significant risk laser device.
  • the laser sources of this device are semiconductor laser arrays that are optically coupled to form a single fiber laser output.
  • the delivery system consists of a single flexible optical fiber. The device delivers continuous wave laser light only.
  • the device is designed specifically to effect microbial cell optical destruction, while preserving and without substantial damage optically or thermally to the human tissue at the infection site being irradiated.
  • the NOVEONTM system was designed to harness the known photo-lethal characteristics of these precise energies to kill pathogenic microorganism at far lower energy levels and heat deposition than is generally necessary to kill pathogens using laser-based thermal sterilization means.
  • topical intra-nasal antimicrobial agents are recognized as the preferred method for preventing (distal-site) infections because of their demonstrated effectiveness and widespread desire to minimize the use of systemic antimicrobials.
  • the design of this protocol includes a number of important factors have been considered. Foremost is the need to assure that the amount of energy used in the Nares is safe for the nasal and nares tissues. Furthermore, significant human and histological tests have been done with the Noveon laser in the areas that the study is treating
  • a cylindrical diffusing optical fiber tip for near infrared light delivery was fabricated specifically for uniform illumination of a length of 1.5 cm, to then be placed in a transparent catheter (of given width) to prevent placement too far anteriorly in the nostril, and guarantee a uniform power density at all tissues proximal to the catheter within the nostril.
  • the tip included an optically transmissive, light diffusing, fiber tip assembly having an entrance aperture through a proximal reflector, a radiation-scattering, transmissive material (e.g. a poly-tetrafluoroethylene tube) surrounding an enclosed cavity (e.g. a cylindrical void filled with air or another substantially non-scattering, transparent medium), and a distal reflective surface.
  • a radiation-scattering, transmissive material e.g. a poly-tetrafluoroethylene tube
  • enclosed cavity e.g. a cylindrical void filled with air or another substantially non-scattering, transparent medium
  • distal reflective surface e.g. a distal reflective surface
  • the scattering medium has a prescribed inner diameter. This inner diameter of the scattering material is designed such that the interaction with this material and the multiple reflections off of the cavity reflectors interact to provide a substantially proscribed axial distribution of laser radiation over the length of the tip apparatus. Suitable choices of tip dimensions provide control over the emitted axial and azimuthal energy distributions.
  • Diode lasers in the near infrared range have a very low absorption coefficient in water; hence, they achieve relatively deep optical penetration in tissues that contain 80% water (such as the dermis, the oral mucosa, bone and the gingiva.
  • the depth of penetration (before photon absorption) of the greatest amount of the incident energy is about 1.5 cm. This allows the near infrared laser energy to pass through water with minimal absorption, producing thermal effects deeper in the tissue and the photons are absorbed by the deeper tissue pigments.
  • This photobiology allows for controlled, deeper soft-tissue irradiation and decontamination, as the photons that emerge from the dispersion tip in a uniform dosimetry from the diffusing tip absorbed by blood and other tissue pigments.
  • This investigational protocol was designed to demonstrate that the Noveon Laser treatment is able to produce reduction in Nasal carriage of MRSA in patients with previously "culture positive" history.
  • This investigational protocol was an open-label study of subjects who are colonized with MRSA in the nares (nostril). The study was done in two parts.
  • the NOVEONTM laser was used for two (2) six-minute treatments in each nostril on day (1) and day (3) of the study.
  • the dosimetries used are shown in the Table 55, below.
  • the laser was calibrated before the first treatment of the day. Intermittent temperature testing of the treatment site was performed on each subject using a noncontact infrared thermometer (Raytek Minitemp), 30-60 second intervals. If a temperature of 110 F degrees was reached, or the patient complained of pain, the laser treatment was interrupted and only resumed when the patient was comfortable. Inturruption only occurred once in 40 treatments (20 nostrils x 2 treatments over three days), and was resumed 30 seconds later to completion.
  • Tables 56-58 represent the mean values of the triplicate CFU counts and plating of each swab from each nostril, pre and post laser therapy (for this data set the mean is the sum of the observed and counted CFU's per plate, divided by the number of counted plates).
  • Table 56 Initial Analysis Pre swab Post-treatment swab left right left right Patient s. aureus Average MRSA Average s. aureus Average MRSA Average s. aureus Average MRSA Average s.
  • a second human study was conducted, to further evaluate the therapeutic potential of the NOVEONTM laser system, including its ability to reverse drug resistance in bacteria.
  • the study was conducted in a similar manner as Part One, above. Outcome measures assessed included both laboratory study and clinical observations.
  • Positive anterior nares cultures were obtained in six patients (12 nostrils) having nasal colonization of MRSA or MSSA, before initiating bacterial photodamage through doses of phototherapeutic near infrared radiation.
  • One patient had MRSA only, 3 had MSSA only, and 2 had both MRSA and MSSA. All MRSA and MSSA were cultured and verified to be resistant to erythromycin.
  • Antimicrobial paste (generic 2% erythromycin) was placed on a cotton tipped swab for application after phototherapeutic near infrared radiation.
  • the swab was inserted approximately 1 cm in to the anterior nares of the subject, rotated 360 degrees several times and removed. The application of erythromycin was maintained for 3 times a day for the remainder of the study.
  • the laser was calibrated before the first treatment of the day and between each patient.
  • the NOVEONTM laser was used for four six-minute treatments of the nares at the following sets of dosimetries (Tables 59), which were evaluated for safety in previous studies.
  • Tables 59 were evaluated for safety in previous studies.
  • each patient underwent exposure with the Noveon for 7 minutes (energy density - 207 J/cm2) to each anterior nostril on Day 1 and on Day 3.
  • the treatment was divided into two parts, an approximately 3-minute exposure using a combination of 870 nm and 930 nm and an approximately 3-minute exposure of 930 nm alone.
  • Temperatures of the nares were recorded every 30 seconds with an IR temperature thermometer.
  • the Erythromycin resistant MRSA was completely cleared by culture in all 3 carriers, as was the E-mycin resistant MSSA in four of the five (5) carriers after the second laser treatment on day 3 and remained clear on day 5.
  • the E-mycin resistant MSSA baseline count > 1000 CFU's
  • No sequelae or adverse events were observed.
  • the average maximum temperature of the nares reached in all patients was 99F.
  • NOVEONTM laser exposure at a non-damaging energy density and approximately physiologic temperatures, re-sensitized erythromycin resistant MRSA and MSSA to 2% generic erythromycin paste. Photodamage to the organism results in sensitivity to antibiotics in otherwise drug resistant strains.
  • the NOVEONTM laser system provides for local reduction of drug resistant microbes and a concomitant reduction of bio-burden in: e.g., wounds, mucosal or cutaneous tissues, and other colonized or infected areas such as surgical sites and tissue/medical device interfaces, which are prone to contamination particularly by nosocomial strains of microbes frequently having multidrug resistance phenotypes.
  • FIG 22 illustrates a schematic diagram of a therapeutic radiation treatment device according one embodiment of the present disclosure.
  • the therapeutic system 110 includes an optical radiation generation device 112, a delivery assembly 114, an application region 116, and a controller 118.
  • the optical radiation generation device includes one or more suitable lasers, L1 and L2.
  • a suitable laser may be selected based on a degree of coherence.
  • a therapeutic system can include at least one diode laser configured and arranged to produce an output in the near infrared region.
  • Suitable diode lasers can include a semiconductor materials for producing radiation in desired wavelength ranges, e.g., 850nm-900nm and 905nm-945nm.
  • Suitable diode laser configurations can include cleave-coupled, distributed feedback, distributed Bragg reflector, vertical cavity surface emitting lasers (VCSELS), etc.
  • the delivery assembly 114 can generate a "flat-top" energy profile for uniform distribution of energy over large areas.
  • a diffuser tip 10 may be included which diffuses treatment light with a uniform cylindrical energy profile in an application region 116 (e.g. a nasal cavity as described in the example above).
  • the optical radiation generation device 112 can include one or more lasers, e.g., laser oscillators L1 and L2.
  • one laser oscillator can be configured to emit optical radiation in a first wavelength range of 850 nm to 900 nm, and the other laser oscillator can be configured to emit radiation in a second wavelength range of 905 nm to 945 nm.
  • one laser oscillator is configured to emit radiation in a first wavelength range of 865 nm to 875 nm
  • the other laser oscillator 28 is configured to emit radiation in a second wavelength range of 925 nm to 935 nm.
  • the geometry or configuration of the individual laser oscillators may be selected as desired, and the selection may be based on the intensity distributions produced by a particular oscillator geometry or configuration.
  • the delivery assembly 114 includes an elongated flexible optical fiber 119 adapted for delivery of the dual wavelength radiation from the oscillators 26 and 28 to diffuser tip 10 to illuminate the application region 116.
  • the delivery assembly 14 may have different formats (e.g., including safety features to prevent thermal damage) based on the application requirements.
  • the delivery assembly 114 or a portion thereof e.g. tip 10) may be constructed with a size and with a shape for inserting into a patient's body.
  • the delivery assembly 114 may be constructed with a conical shape for emitting radiation in a diverging-conical manner to apply the radiation to a relatively large area.
  • Hollow waveguides may be used for the delivery assembly 114 in certain embodiments.
  • Other size and shapes of the delivery assembly 14 may also be employed based on the requirements of the application site.
  • the delivery assembly 114 can be configured for free space or free beam application of the optical radiation, e.g., making use of available transmission through tissue at NIMELS wavelengths described herein. For example, at 930nm (and to a similar degree, 870nm), the applied optical radiation can penetrate patient tissue by up to 1 cm or more. Such embodiments may be particularly well suited for use with in vivo medical devices as described herein.
  • delivery assembly 114 may terminate in a delivery head of the type described in detail below.
  • the delivery head may connect to a receptacle used for positioning the head near a target region on the body part of a subject.
  • the delivery head may communicate with controller 118 e.g., via a wired, wireless or other suitable link.
  • the deliver head and/or receptacle may include one or more sensors in communication with controller 118,
  • the controller 118 includes a power limiter 124 connected to the laser oscillators L1 and L2 for controlling the dosage of the radiation transmitted through the application region 116, such that the time integral of the power density of the transmitted radiation per unit area is below a predetermined threshold, which is set up to prevent damages to the healthy tissue at the application site.
  • the controller 118 may further include a memory 126 for storing treatment information of patients.
  • the stored information of a particular patient may include, but not limited to, dosage of radiation, (for example, including which wavelength, power density, treatment time, skin pigmentation parameters, etc.) and application site information (for example, including type of treatment site (lesion, cancer, etc.), size, depth, etc.).
  • controller 118 may communicate with one or more sensors of any suitable type, e.g. a temperature sensor which monitors the temperature of the target region 116. Base don information from the sensor, the controller 118 may change one or more of the parameters (e.g. power, power desity, energy desity, pulse rate, etc.) of the applied therapeutic light. For example, in some emdodiments controller 118 shuts off treatment light if a temperature sensor senses that the temperature of the treatment area is greater than a threshold value
  • the memory 126 may also be used to store information of different types of diseases and the treatment profile, for example, the pattern of the radiation and the dosage of the radiation, associated with a particular type of disease.
  • the controller 118 may further include a dosimetry calculator 128 to calculate the dosage needed for a particular patient based on the application type and other application site information input into the controller by a physician.
  • the controller 118 further includes an imaging system for imaging the application site. The imaging system gathers application site information based on the images of the application site and transfers the gathered information to the dosimetry calculator 128 for dosage calculation. A physician also can manually calculate and input information gathered from the images to the controller 118.
  • the controller may further include a control panel 130 through which, a physician can control the therapeutic system manually.
  • the therapeutic system 10 also can be controlled by a computer, which has a control platform, for example, a WINDOWS TM based platform.
  • the parameters such as pulse intensity, pulse width, pulse repetition rate of the optical radiation can be controlled through both the computer and the control panel 30.
  • Figures 23a-23d show different temporal patterns of the optical radiation that can be delivered from the therapeutic system to the application site.
  • the optical radiation can be delivered in one wavelength range only, for example, in the first wavelength range of 850 nm to 900 nm, or in the range of 865 nm to 875 nm, or in the second wavelength range of 905 nm to 945 nm, or in the range of 925 nm to 935 nm, as shown in Figure 23a .
  • the radiation in the first wavelength range and the radiation in the second wavelength range also can be multiplexed by a multiplex system installed in the optical radiation generation device 112 and delivered to the application site in a multiplexed form, as shown in Figure 23b .
  • the radiation in the first wavelength range and the radiation in the second wavelength range can be applied to the application site simultaneously without passing through a multiplex system.
  • Figure 23c shows that the optical radiation can be delivered in an intermission-alternating manner, for example, a first pulse in the first wavelength range, a second pulse in the second wavelength range, a third pulse in the first wavelength range again, and a fourth pulse in the second wavelength range again, and so on.
  • the interval can be CW (Continuous Wave), one pulse as shown in Figure 18c , or two or more pulses (not shown).
  • Figure 23d shows another pattern in which the application site is first treated by radiation in one of the two wavelength ranges, for example, the first wavelength range, and then treated by radiation in the other wavelength range.
  • the treatment pattern can be determined by the physician based on the type, and other information of the application site.
  • FIG 24A illustrates an exemplary receptacle 1030a.
  • the receptacle 1030a includes connection mechanisms 1032a and 1034a, a positioning region 1036a, and a bottom region 1038a.
  • the receptacle 1030a can receive a therapeutic output head (e.g., laser output, optical output, heat output, sonic output, etc.), which is connected to a therapeutic device that delivers the therapeutic output to the therapeutic output head.
  • the connection mechanisms 1032a and 1034a interlock the receptacle 1030a and the therapeutic output head.
  • the positioning region 1036a enables the receptacle 1030a to be positioned above a body part (e.g., digit, skin, nail, hair, etc.).
  • the receptacle 1030a can be, for example, utilized with the therapeutic system 110 of figure 22 .
  • the receptacle 1030a can receive the the delivery assembly 114 for delivery of the radiation to the patient.
  • the receptacle 1030a includes a bottom wall and four vertical walls enabling the therapeutic output head to be placed completely or partially within the receptacle 1030a.
  • the bottom wall and the four walls can be interconnected together along the seams (e.g., welded, bonded, etc.) to form a hollow receptacle with an open top.
  • the receptacle 1030a is form molded plastic.
  • the receptacle 1030a is constructed from plastic, metal, and/or any other type of material.
  • the receptacle 1030a can be, for example, stiff and/or flexible.
  • the walls of the receptacle 1030a are constructed from a flexible silicon and the bottom wall is constructed from a stiff plastic.
  • connection mechanisms 1032a and 1034a include a single use interlock mechanism, an interlocking releasable mechanism (e.g., releasable clip, releasable tab, etc.), guide mechanism, alignment mechanism, and/or any other type of connection component.
  • the receptacle 1030a positions the therapeutic output head at a predetermined distance above the body part (e.g., the connection mechanisms 1032a and 1034a position the therapeutic output head at the predetermined distance).
  • the predetermined distance can be an optimal distance for treatment of a specified disease and/or condition on the body part.
  • the predetermined distance is two centimeters between the laser output from the therapeutic output head and a toenail. In this example, the ten centimeters enables the laser output to diffuse while still retaining energy for destroying microbes on the toenail.
  • the positioning of the therapeutic output head at the predetermined distance above the body part can enable the therapeutic output head to remain sterile or nearly sterile while the receptacle 1030a is in contact with the body part.
  • the bottom wall of the receptacle 1030a is adjacent to the body part, and the therapeutic output head does not contact the body part.
  • the positioning region 1036a is light transparent.
  • the positioning region 1036a can enable any light wavelength or range of light wavelengths (e.g., near infrared range, near visual range, etc.) to pass through the bottom of the receptacle 1030a via the positioning region 1036a.
  • the positioning region 1036a is light transparent enabling fight wavelengths 870 and 930 nanometers (nm) to pass through the bottom of the receptacle 1030a via the positioning region 1036a.
  • the bottom region 1038a is light opaque or nearly light opaque.
  • the bottom region 1038a can prevent, nearly prevent, or reduce any light wavelength or range of light wavelengths from passing through the bottom of the receptacle 1030a.
  • the bottom region 1038a prevents light wavelengths 870 and 930 nm from passing through the bottom of the receptacle 1030a.
  • FIG 24B illustrates a top view of an exemplary receptacle 1030b.
  • the receptacle 1030b includes a positioning region 1036b and a microchip 1038b (e.g., storage device, computer readable storage device, memory, etc.).
  • the positioning region 1036b is positioned on a bottom of the receptacle 1030b to enable an output from the therapeutic output head to pass through the receptacle 1030b via the positioning region 1036b.
  • the microchip 1038b can include or be coupled to a sensor (e.g., temperature sensor, humidity sensor, position sensor, pressure sensor, accelerometer, etc.) and/or an identification mechanism (e.g., encryption mechanism, authentication mechanism, single use mechanism, etc).
  • the microchip 1038b can be positioned to interface with the therapeutic output head. In other words, in this example, the microchip 1038b and the therapeutic output head are interconnected (e.g., electrically, physically, etc.).
  • the identification mechanism enables the receptacle 1030b to only be utilized once.
  • the receptacle 1030b is a one-time use medical apparatus that is disposed of after use on the body part.
  • the microchip 1038b can include a destruction mechanism that self-destructs after a specified time period of use (e.g., ten seconds, twenty seconds, etc.).
  • the destruction mechanism destroys the identification mechanism and the destruction of the identification mechanism prevents the receptacle 1030b from being re-used since the therapeutic output head cannot verify the identity of the receptacle 1030b.
  • the microchip 1038b includes an encryption mechanism that enables authentication of the receptacle 1030b with the therapeutic output head.
  • the therapeutic output head and/or the therapeutic device can query the encryption mechanism to determine the identify of the receptacle 1030b.
  • the encryption mechanism can respond with an authentication response (e.g., key, signature, security token, etc.).
  • the therapeutic output head and/or the therapeutic device can process the authentication response to verify that the receptacle 1030b is valid and/or other information associated with the receptacle 1030b (e.g., that the receptacle 1030b can operate with the therapeutic output head, therapeutic parameters of the receptacle 1030b, the serial number of the receptacle 1030b, the manufacturer of the receptacle 1030b, etc.). If the receptacle 1030b is not validated, the therapeutic output head and/or the therapeutic device can automatically de-activate until a validated receptacle 1030b is connected to the therapeutic output head.
  • the authentication mechanism for the receptacle 1030b can advantageously protect patients by ensuring that the bodies are not re-used, i.e., the safety of the patient is increased.
  • the microchip 1038b includes a temperature sensor.
  • the temperature sensor can sense the temperature of the bottom side of the receptacle 1030b.
  • the temperature sensor can deactivate the therapeutic output based on the temperature (e.g., outside of a set temperature range, drops below a minimum temperature, etc.).
  • the temperature sensor can communicate the temperature data to the therapeutic output head and/or the therapeutic device, and the therapeutic output head and/or the therapeutic device can deactivate or otherwise modify the therapeutic output based on the temperature (e.g., below a minimum temperature, above a maximum temperature, etc.).
  • the temperature sensor can advantageously protect patients by ensuring that the therapeutic output is deactivated if the body part becomes too hot (i.e., temperature is above a maximum temperature) or if the receptacle 1030b is removed from the body part (i.e., temperature drops below a minimum temperature).
  • each receptacle 1030b is uniquely identified (e.g., unique serial number, etc.) and/or the microchip 1038b includes the unique identification information.
  • each set of one or more bodies is uniquely identified (e.g., each size and lot includes a unique lot number, etc.) and/or the microchip 1038b includes the identification information.
  • the microchip 1038b includes information associated with the therapeutic output (e.g., pulse, wavelength, time, diameter of target, etc.).
  • the microchip 1038b can be embedded into a side of the receptacle 1030b (e.g., the microchip 1038b is a radio frequency identification (RFID) device).
  • RFID radio frequency identification
  • FIG. 10B illustrates the microchip 1038b
  • the receptacle 1030b can include any type of device and/or mechanism that can include identification information associated with the receptacle 1030b (e.g., barcode, passive radio device, read-only memory, etc.).
  • FIG. 25 illustrates an exemplary therapeutic output system 1100.
  • the therapeutic output system 1100 includes a therapeutic device (not shown), a therapeutic output head 1120, and a receptacle 1130 for positioning the therapeutic output head 1120 on a body part (e.g., over a toenail, over a fingernail, etc.).
  • the therapeutic output head 1120 includes a cable 1122, connection mechanisms 1124 and 1128, a microchip interface 1126, and an output connector 1129.
  • the cable 1122 transmits the output between the therapeutic output head 1120 and the therapeutic device.
  • the connection mechanisms 1124 and 1128 connect and/or lock the therapeutic output head 1120 with/to the receptacle 1130.
  • the microchip interface 1126 connects and/or communications with a microchip on the receptacle 1130 (e.g., electronically connection, radio connection, optical connection, etc.).
  • the output connector 1129 transmits the therapeutic output from the therapeutic output head 1120 to the body part (e.g., light at a specified wavelength to an area of skin on the patient's arm).
  • the receptacle 1130 includes connection mechanisms 1132 and 1134 and a positioning region 1136.
  • the connection mechanism 1132 of the receptacle 1130 interfaces with the connection mechanism 1124 of the therapeutic output head 1120 to connect the receptacle 1130 to the therapeutic output head 1120 (e.g., slide mechanism).
  • the connection mechanism 1134 of the receptacle 1130 interfaces with the connection mechanism 1128 of the therapeutic output head 1120 to connect the receptacle 1130 to the therapeutic output head 1120 (e.g., tab lock mechanism).
  • the positioning region 1136 is transparent and enables the therapeutic output to pass through to the body part.
  • the therapeutic output head 1120 connects with the receptacle 1130.
  • the connection between the therapeutic output head 1120 and the receptacle 1130 can be temporary and quickly reversed for ease of use of the therapeutic output system 1100.
  • Figure 26A illustrates a bottom view of an exemplary receptacle 1230a.
  • the receptacle 1230a includes a positioning region 1236a and a fixing mechanism 1242a.
  • the fixing mechanism 1242a can be utilized to affix the receptacle 1230a to a patient's body part (e.g., via an adhesive pad, via a clamping mechanism, via glue, etc).
  • the fixing mechanism 1242a is a adhesive pad that sticks the receptacle 1230a to the patient's toe.
  • FIG 26A illustrates the fixing mechanism 1242a shaped as a butterfly
  • the fixing mechanism 1242a can be any shape and/or form (e.g., rectangular, elliptical, half-moons, etc.) and/or can utilize any type of mechanism to affix the receptacle 1230a to the patient's body part.
  • Figure 26B illustrates a bottom view of an exemplary receptacle 1230b.
  • the receptacle 1230b includes a positioning region 1236b and a fixing mechanism 1242b.
  • the fixing mechanism 1242b can be utilized to affix the receptacle 1230b to a patient's body part (e.g., via an adhesive pad, etc.).
  • Figure 26C illustrates a bottom view of an exemplary receptacle 1230c.
  • the receptacle 1230c includes a positioning region 1236c and a fixing mechanism 1242c, 1243c, 1244c, and 1245c.
  • the fixing mechanism can include straps 1242c and 1244c and connection mechanism 1243c and 1245c, respectively.
  • the straps 1242c and 1244c can wrap around a patient's body part and the connection mechanism 1243c and 1245c can engage to affix the receptacle 1230c to the patient's body part.
  • Figure 27A illustrates a bottom view of an exemplary receptacle 1330a.
  • the receptacle 1330a includes a positioning region 1336a.
  • the positioning region 1336a is transparent for a specified wavelength of the therapeutic output and is semi-transparent for visible light wavelengths.
  • Figure 27B illustrates a bottom view of an exemplary receptacle 1330b.
  • the receptacle 1330b includes a positioning region 1336b.
  • the positioning region 1336b includes an indicia to position the receptacle 1330b on a patient's body part.
  • FIG. 13B illustrates a fine crosshair
  • the positioning region 1336b can include any type of crosshair (e.g., duplex crosshair, target crosshair, etc.) or other indicia may be used.
  • Figure 27C illustrates a bottom view of an exemplary receptacle 1330c.
  • the receptacle 1330c includes a positioning region 1336c.
  • the positioning region 1336c is rectangular and enables the receptacle 1330c to be positioned on a patient's body part.
  • the therapeutic output can be, for example, shaped to be rectangular.
  • Figure 27D illustrates a bottom view of an exemplary receptacle 1330d.
  • the receptacle 1330d includes a positioning region 1336d.
  • the positioning region 1336d includes double circles to position the receptacle 1330d on a patient's body part.
  • FIG. 28 illustrates an exemplary therapeutic output system 1400.
  • the therapeutic output system receptacle 1400 includes a therapeutic device 1440 (e.g., laser output device, infrared heat output device, etc.) and interconnected therapeutic output head and receptacle devices 1430a, 1430b, 1430c, and 1430d.
  • therapeutic device 1440 may include a NIMELS antimicrobial system of the type described above.
  • the interconnected therapeutic output head and receptacle devices 1430a, 1430b, 1430c, and 1430d are affixed to toes 1422a, 1422b, 1422c, and 1422d, respectively, on a patient's foot 1420.
  • the receptacles are packaged in sets for use with the therapeutic output system 1400 (e.g., three sterile receptacles, five sterile receptacles, etc.).
  • the bodies are packaged as two large receptacles and two small receptacles.
  • the bodies are packaged as one large receptacle and four small receptacles.
  • Figure 29A illustrates a side view of an exemplary receptacle 1530a.
  • the receptacle 1530a includes connection mechanisms 1534a, a fixing mechanism 1542a, and is defined by side walls 1532a.
  • the receptacle 1530a can receive a therapeutic output head.
  • the connection mechanisms 1534a interlock the receptacle 1530a and the therapeutic output head (in this example, a tab lock mechanism).
  • the receptacle 1530a is positioned over a body part 1560a (in this example, a toe) and aligned over a body area for treatment 1562a (in this example, a toenail).
  • the fixing mechanism 1542a affixes the receptacle 1530a to the body part 1560a (in this example, the adhesive wings are affixed to the body part 1560a).
  • Figure 29B illustrates another side view of an exemplary receptacle 1530b.
  • the receptacle 1530b includes a connection mechanism 1536b, a fixing mechanism 1542b, cooling vents 1538b, and is defined by side walls 1533b.
  • the receptacle 1530b can receive a therapeutic output head.
  • the connection mechanisms 1536b interlock the receptacle 1530b and the therapeutic output head (in this example, a feed mechanism).
  • the receptacle 1530b is positioned over a body part and aligned over a body area for treatment.
  • the fixing mechanism 1542b affixes the receptacle 1530b to the body part.
  • the cooling vents 1538b enable air circulation to cool the therapeutic output head (e.g., to provide overheating of the therapeutic output head, etc.).
  • Figure 29C illustrates a top view of an exemplary receptacle 1530c.
  • the receptacle 1530c includes connection mechanisms 1534c, a fixing mechanism 1542c, a microchip 1531c, and a positioning region 1533c.
  • the receptacle 1530c can receive a therapeutic output head.
  • the connection mechanism 1534c interlock the receptacle 1530c and the therapeutic output head (in this example, a tab lock mechanism).
  • the receptacle 1530c is positioned over a body part and aligned over a body area for treatment utilizing the positioning region 1533c.
  • the fixing mechanism 1542c affixes the receptacle to the body part.
  • the microchip 1531c provides identification and/or authorization of the receptacle 1530c to the therapeutic output head.
  • Figure 29D illustrates a perspective view of an exemplary receptacle 1530d.
  • the receptacle 1530d includes a connection mechanism 1534d, a connection mechanism 1536d, a microchip 1531d, cooling vents 1538d, a fixing mechanisms 1544d, and a fixing mechanism protection cover 1546d.
  • the receptacle 1530d can receive a therapeutic output head.
  • the connection mechanisms 1534d and 1536d interlock the receptacle 1530d and the therapeutic output head.
  • the receptacle 1530d is positioned over a body part and aligned over a body area for treatment utilizing a positioning region (not shown).
  • the fixing mechanism 1544d affixes the receptacle 1530d to the body part.
  • the fixing mechanism 1544d is protected via the fixing mechanism protection cover 1546d (e.g., protects the adhesive aspects of the fixing mechanism 1544d from contamination, etc.).
  • the microchip 1531d provides identification and/or authorization of the receptacle 1530c to the therapeutic output head.
  • the cooling vents 1538d enable air circulation to cool the therapeutic output head.
  • Figure 30A illustrates a side view of an exemplary receptacle 1630a.
  • the receptacle 1630a includes connection mechanisms 1634a, a fixing mechanism 1642a, and is defined by side walls 1632a.
  • the receptacle 1630a can receive a therapeutic output head.
  • the connection mechanisms 1634a interlocks the receptacle 1630a and the therapeutic output head (in this example, a tab lock mechanism).
  • the receptacle 1630a is positioned over a body part and aligned over a body area for treatment.
  • the fixing mechanism 1642a affixes the receptacle 1630a to the body part.
  • Figure 30B illustrates another side view of an exemplary receptacle 1630b.
  • the receptacle 1630b includes a connection mechanism 1636b, a fixing mechanism 1642b, cooling vents 1638b, and is defined by side walls 1633b.
  • the receptacle 1630b can receive a therapeutic output head.
  • the connection mechanisms 1636b interlocks the receptacle 1630b and the therapeutic output head (in this example, a feed mechanism).
  • the receptacle 1630b is positioned over a body part and aligned over a body area for treatment.
  • the fixing mechanism 1642b affixes the receptacle 1630b to the body part.
  • the cooling vents 1638b enable air circulation to cool the therapeutic output head (e.g., to provide overheating of the therapeutic output head, etc.).
  • Figure 30C illustrates a top view of an exemplary receptacle 1630c.
  • the receptacle 1630c includes connection mechanisms 1634c, a fixing mechanism 1642c, a microchip 1631c, and a positioning region 1633c.
  • the receptacle 1630c can receive a therapeutic output head.
  • the connection mechanism 1634c interlocks the receptacle 1630c and the therapeutic output head (in this example, a tab lock mechanism).
  • the receptacle 1630c is positioned over a body part and aligned over a body area for treatment utilizing the positioning region 1633c.
  • the fixing mechanism 1642c affixes the receptacle 1630c to the body part.
  • the microchip 1631c provides identification and/or authorization of the receptacle 1630c to the therapeutic output head.
  • Figure 30D illustrates a perspective view of an exemplary receptacle 1630d.
  • the receptacle 1630d includes a connection mechanism 1634d, a connection mechanism 1636d, a microchip 1631d, cooling vents 1638d, a fixing mechanism 1644d, and a fixing mechanism protection cover 1646d.
  • the receptacle 1630d can receive a therapeutic output head.
  • the connection mechanisms 1634d and 1636d interlock the receptacle 1630d and the therapeutic output head.
  • the receptacle 1630d is positioned over a body part and aligned over a body area for treatment utilizing a positioning region (not shown).
  • the fixing mechanism 1644d affixes the receptacle 1630d to the body part.
  • the fixing mechanism 1644d is protected via the fixing mechanism protection cover 1646d (e.g., protects the adhesive aspects of the fixing mechanism 1644d from contamination, etc.).
  • the microchip 1631d provides identification and/or authorization of the receptacle 1630c to the therapeutic output head.
  • the cooling vents 1638d enable air circulation to cool the therapeutic output head.
  • FIG 31 illustrates an exemplary therapeutic output system 1700.
  • the therapeutic output system 1700 includes a therapeutic output head 1720 and a receptacle 1730.
  • the receptacle 1730 includes mechanisms to connect with the therapeutic output head 1720.
  • FIG 32 illustrates an exemplary therapeutic output head 1820.
  • the therapeutic output head 1820 includes a fiber optic distal end 1822, a thermopile sensor 1824, and a connection mechanism 1826.
  • the fiber optic distal end 1822 delivers an output (e.g., laser output, infrared output, etc.) to the therapeutic output head 1820 from a therapeutic device via a fiber optic cable.
  • the thermopile sensor 1824 monitors the temperature of the output.
  • the connection mechanism 1826 connects the therapeutic output head 1820 with a receptacle (e.g., the receptacle 1730 of Figure 31 ).
  • Figure 33 is a flowchart 1900 of an exemplary therapeutic output process utilizing, for example, the therapeutic output system 1400 of figure 28 .
  • a healthcare provider and/or a patient connects (1910) the receptacle with a therapeutic output head to form the interconnected therapeutic output head and receptacle device 1430a.
  • the healthcare provider and/or the patient positions (1920) the device 1430a on a body part (e.g., a toenail).
  • the healthcare provider and/or the patient secures (1930) the device 1430a to the body part via a fixing mechanism (e.g., an adhesive pad).
  • the healthcare provider and/or the patient sets (1940) one or more output parameters on the therapeutic device 1440.
  • the healthcare provider and/or the patient turns on (1950) the therapeutic device 1440 for a specified time period (e.g., one minute, three minute, etc.). After the cycle for the therapeutic device 1440 is complete (i.e., the therapeutic device 1440 turns off), the healthcare provider and/or the patient removes (1960) the therapeutic output head from the receptacle. The healthcare provider and/or the patient disposes (1970) of the receptacle (e.g., medical waste, trash, etc.).
  • a specified time period e.g., one minute, three minute, etc.
  • any of the receptacles described herein can be, for example, utilized with the therapeutic system 110 of figure 22 .
  • the receptacle can receive the the delivery assembly 114 for delivery of the radiation to the patient.
  • the phrases “light”, “optical”, etc. are not limited to the visible spectrum, but may refer to electromagnetic radiation at any wavelength including, e.g., the infrared.

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Claims (15)

  1. Vorrichtung (110, 1100, 1400, 1700) zum Positionieren eines Lichtzuführungskopfs (1120, 1720, 1820) einer therapeutischen Behandlungsvorrichtung (1440) in Nähe zu einem Körper mit einem Behandlungszielbereich, wobei die Vorrichtung (110, 1100, 1400, 1700) aufweist:
    eine Positioniervorrichtung, aufweisend:
    eine Aufnahme (1330a), die ein mindestens teilweise umschlossenes Volumen definiert, um mindestens einen Abschnitt des Zuführungskopfs (1120, 1720, 1820) aufzunehmen, wobei die Aufnahme eine Behandlungszuführungsfläche hat, aufweisend:
    einen lichtaussendenden Bereich (1336a), der zumindest teilweise lichtdurchlässig ist für therapeutisches Licht von dem Behandlungskopf, wobei der lichtaussendende Bereich lichtdurchlässig ist für therapeutisches Licht mit einer Wellenlänge von 870 nm oder 930 nm und halbdurchlässig für Licht von Wellenlängen im sichtbaren Bereich, und einen lichtabschirmenden Bereich, der relativ weniger lichtdurchlässig für das therapeutische Licht ist als der lichtaussendende Bereich;
    eine Fixierungseinrichtung, die die Aufnahme so an dem Körperteil befestigt, dass der lichtaussendende Bereich der Behandlungszuführungsfläche an den Behandlungszielbereich angrenzt;
    einen digitalen Speicher (126); und
    eine Datenübertragungsverbindung, die so ausgelegt ist, dass sie den Speicher (126) selektiv mit der Behandlungsvorrichtung (1440) verbindet.
  2. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 1, wobei der digitale Speicher (126) von der Behandlungsvorrichtung (1440) über die Datenübertragungsverbindung lesbar und beschreibbar ist, und/oder optional, wobei der digitale Speicher (126) Informationen speichert, die für die Identität der Positioniervorrichtung indikativ sind, und/oder optional, wobei die für die Identität der Positioniervorrichtung indikativen Informationen verschlüsselt sind.
  3. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 2, wobei der digitale Speicher (126) Informationen speichert, die für die Verwendungschronik der Positioniervorrichtung indikativ sind, und/oder optional, wobei die für die Verwendungschronik der Positioniervorrichtung indikativen Informationen verschlüsselt sind.
  4. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 1, ferner aufweisend mindestens einen Sensor, und wobei die Datenübertragungsverbindung so ausgelegt ist, dass sie selektiv den mindestens einen Sensor mit der Behandlungsvorrichtung (1440) verbindet, und/oder optional, wobei der mindestens eine Sensor einen Temperatursensor aufweist, oder optional, wobei der mindestens eine Sensor mindestens einen aus der Liste ausgewählten aufweist, bestehend aus: Positionssensor, Feuchtigkeitssensor, Positionssensor, Drucksensor, Beschleunigungssensor, Fotodetektor, Brechkraftsensor und Lichtwellenlängensensor.
  5. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 1, wobei die Datenübertragungsverbindung den Speicher (126) nur dann mit der Behandlungsvorrichtung (1440) verbindet, wenn der Zuführungskopf (1120, 1720, 1820) von der Aufnahme aufgenommen wird, oder optional, wobei die Datenübertragungsverbindung mindestens eine aus der Liste aufweist, bestehend aus: einer elektrischen Verbindung, einer verkabelten Verbindung, einer kabellosen Verbindung, einer Funkverbindung, einer optischen Datenübertragungsverbindung und einer induktiven Verbindung.
  6. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 1, ferner aufweisend einen Steckverbinder, der den Zuführungskopf (1120, 1720, 1820) mit der Aufnahme in einer gewünschten Ausrichtung verbindet, und /oder optional, wobei der Steckverbinder so ausgelegt ist, dass er die Verbindung des Zuführungskopfs (1120, 1720, 1820) mit der Aufnahme in anderen Ausrichtungen als der gewünschten Ausrichtung verhindert.
  7. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 1, aufweisend mindestens ein Zeichen zum Erleichtern der Ausrichtung des lichtaussendenden Bereichs auf den Zielbereich.
  8. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 1, wobei der lichtabschirmende Bereich am Rand des lichtaussendenden Bereichs liegt.
  9. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 1, wobei die Fixierungseinrichtung ein in Kontakt mit einem Abschnitt der Behandlungszuführungsfläche anhaftendes Material aufweist, und/oder optional, wobei sich mindestens ein Abschnitt des anhaftenden Materials über die Behandlungszuführungsfläche hinaus erstreckt.
  10. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 1, wobei die Aufnahme mindestens einen Anschluss aufweist, der eine Fluidverbindung zwischen dem mindestens teilweise umschlossenen Volumen und dem Äußeren der Aufnahme bereitstellt.
  11. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 1, aufweisend einen Mikrochip, der den digitalen Speicher (126) aufweist.
  12. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 1, wobei der lichtaussendende Bereich ein streuendes Element aufweist, das zumindest teilweise therapeutisches Licht von dem Zuführungskopf (1120, 1720, 1820) auf den Zielbereich streut, oder optional, wobei der lichtaussendende Bereich zumindest teilweise lichtdurchlässig ist für Licht im Nah-Infrarot.
  13. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 1, ferner aufweisend die Behandlungsvorrichtung (1440), und wobei die Behandlungsvorrichtung (1440) einen Controller aufweist; der Controller ist so ausgelegt, dass er Informationen, die in dem digitalen Speicher (126) gespeichert sind, über die Datenübertragung und Steuerung der Zuführung von Behandlungslicht auf der Grundlage der Informationen empfängt, und/oder optional, wobei die Informationen, die in dem digitalen Speicher (126) gespeichert sind, Informationen aufweisen, die indikativ für die Identität der Positioniervorrichtung oder die Verwendungschronik der Positioniervorrichtung sind; und der Controller ist so konfiguriert, dass er die Zuführung von Behandlungslicht auf der Grundlage der Identität oder der Verwendungschronik der Positioniervorrichtung steuert, oder optional, wobei die Informationen, die in dem digitalen Speicher (126) gespeichert sind, Informationen aufweisen, die indikativ für die Verwendungschronik der Positioniervorrichtung sind, und wobei der Controller so ausgelegt ist, dass er eine Zuführung von Behandlungslicht unterbindet, wenn eine vorherige Verwendung der Positioniervorrichtung angegeben wird, und/oder optional, wobei die Informationen, die in dem digitalen Speicher (126) gespeichert sind, verschlüsselte Informationen enthalten und der Controller so ausgelegt ist, dass er die verschlüsselten Informationen entschlüsselt.
  14. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 13, wobei der Controller so ausgelegt ist, dass er Informationen zum Speichern über die Datenübertragungsverbindung in den digitalen Speicher (126) schreibt, oder optional, wobei die Informationen, die zum Speichern in den digitalen Speicher (126) geschrieben wurden, Informationen aufweisen, die indikativ für die Verwendungschronik der Positioniervorrichtung sind.
  15. Vorrichtung (110, 1100, 1400, 1700) nach Anspruch 13 oder Anspruch 14, wobei die Behandlungsvorrichtung (1440) aufweist: eine therapeutische Lichtquelle, die so ausgelegt ist, dass sie Behandlungslicht erzeugt; und eine Zuführungsanordnung, die das Behandlungslicht zu dem Zuführungskopf (1120, 1720, 1820) zuführt; wobei der Controller betriebsfähig mit der Lichtquelle verbunden ist und die Lichtquelle so steuert, dass sie das Behandlungslicht in dem Zielbereich im Wesentlichen in einem ersten Wellenlängenbereich von ungefähr 865 nm bis ungefähr 875 nm oder in einem zweiten Strahlungsbereich mit einer Wellenlänge von ungefähr 925 nm bis ungefähr 935 nm oder beide Wellenlängenbereiche in einer Dosimetrie bereitstellt mit einer Leistungsdichte von ungefähr 0,5 W/cm2 bis ungefähr 40 W/cm2 und einer Energiedichte von ungefähr 200 J/cm2 bis ungefähr 700 J/cm2 und einer Zeitdauer von ungefähr 50 bis ungefähr 720 Sekunden, und wobei das Behandlungslicht dem Zielbereich von dem Zuführungskopf (1120, 1720, 1820) über den lichtaussendenden Bereich der Behandlungszuführungsfläche der Positioniervorrichtung zugeführt wird, und/oder optional, wobei der Controller betriebsfähig mit der Lichtquelle verbunden ist und die Lichtquelle so steuert, dass sie das Behandlungslicht am Zielbereich im Wesentlichen in einem ersten Wellenlängenbereich von ungefähr 865 nm bis ungefähr 875 nm oder in einem zweiten Wellenlängenbereich mit einer Wellenlänge von ungefähr 925 nm bis ungefähr 935 nm bereitstellt und mit einer Dosimetrie mit einer Leistungsdichte von ungefähr 0,5 W/cm2 bis ungefähr 5 W/cm2 und einer Energiedichte von ungefähr 200 J/cm2 bis ungefähr 700 J/cm2, und/oder optional, wobei das Behandlungslicht am Zielbereich eine Punktgröße von mindestens 1 cm hat.
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EP2995346B1 (de) 2019-03-20
US20150157876A1 (en) 2015-06-11
US20110295343A1 (en) 2011-12-01
EP2995346A1 (de) 2016-03-16
EP2349479A4 (de) 2012-04-18
WO2010056537A1 (en) 2010-05-20

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